Anti-GAPDH 抗体 [6C5] - Loading Control

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

ab8245 staining GAPDH in SV40LT-SMC (Rat SV40-transfected aorta smooth cell line) cells.

The cells were fixed with 4% formaldehyde (10 minutes) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5μg/ml and ab202272 at 1/250 dilution overnight at +4°C followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

ab8245 staining GAPDH in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

The cells were fixed with 100% methanol (5 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5 μg/ml and ab6046 at 1 μg/ml overnight at +4°C followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) at 2 μg/ml (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.

Negative controls : 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

ab8245 staining GAPDH in NIH/3T3 (Mouse embryo fibroblast cell line) cells.

The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 1 μg/ml and ab202272 at 1/250 dilution overnight at +4°C followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-B4GALNT1 antibody [EPR28064-64] ab314130 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 70 kDa in Wild-type A549 cell lysates with no signal observed at this size in B4GALNT1 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-B4GALNT1 antibody [EPR28064-64] (<a href='/products/primary-antibodies/b4galnt1-antibody-epr28064-64-ab314130'>ab314130</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

B4GALNT1 knockout A549 cell lysates at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

HEK-293T cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 75 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-p73 antibody [EP436Y] ab40658 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 75 kDa in Wild-type HAP1 cell lysates with no signal observed at this size in TP73 knockout HAP1 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-p73 antibody [EP436Y] (<a href='/products/primary-antibodies/p73-antibody-ep436y-ab40658'>ab40658</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 cell lysates at 20 µg

Lane 2:

TP73 knockout HAP1 cell lysates at 20 µg

Lane 3:

HEK-293 cell lysates at 20 µg

Lane 4:

HeLa cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 69 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-Sortilin/NT3 antibody [EPR23093-58] ab263864 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 100 kDa in Wild-type HeLa cell lysates with no signal observed at this size in SORT1 knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Sortilin/NT3 antibody [EPR23093-58] (<a href='/products/primary-antibodies/sortilin-nt3-antibody-epr23093-58-ab263864'>ab263864</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysates at 20 µg

Lane 2:

Western blot - Human SORT1 (Sortilin/NT3) knockout HeLa cell line (<a href='/products/cell-lines/human-sort1-sortilin-nt3-knockout-hela-cell-line-ab264772'>ab264772</a>) at 20 µg

Lane 3:

SW480 cell lysates at 20 µg

Lane 4:

SH-SY5Y cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 100 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-STAT1 antibody [EPRR21057-168] ab210524 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 87 kDa in Wild-type HeLa cell lysates with no signal observed at this size in STAT1 knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-STAT1 antibody [EPRR21057-168] (<a href='/products/primary-antibodies/stat1-antibody-eprr21057-168-ab210524'>ab210524</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysates at 20 µg

Lane 2:

Western blot - Human STAT1 knockout HeLa cell line (<a href='/products/cell-lines/human-stat1-knockout-hela-cell-line-ab255346'>ab255346</a>) at 20 µg

Lane 3:

A-431 cell lysates at 20 µg

Lane 4:

K562 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 87 kDa

Observed band size: 87 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-NF-kB p65 antibody [EP2161Y] ab76311 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 60 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in RELA knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-NF-kB p65 antibody [EP2161Y] (<a href='/products/primary-antibodies/nf-kb-p65-antibody-ep2161y-ab76311'>ab76311</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 cell lysates at 20 µg

Lane 2:

Western blot - Human RELA knockout MCF7 cell line (ab287740) at 40 µg

Lane 3:

PANC-1 cell lysates at 20 µg

Lane 4:

Saos-2 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 60 kDa

Observed band size: 60 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-OCRL antibody [EP10256] ab181039 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 104 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in OCRL knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-OCRL antibody [EP10256] (<a href='/products/primary-antibodies/ocrl-antibody-ep10256-ab181039'>ab181039</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG cell lysates at 20 µg

Lane 2:

OCRL knockout U-87 MG cell lysates at 20 µg

Lane 3:

PC-3 cell lysates at 20 µg

Lane 4:

THP-1 cell lysates at 20 µg

Lane 5:

Raji cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 104 kDa

Observed band size: 104 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-YTHDF3 antibody [HL2235] ab317173 staining at 1/2000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 70 kDa in Wild-type PC-3 cell lysates with no signal observed at this size in YTHDF3 knockout PC-3 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-YTHDF3 antibody [HL2235] (<a href='/products/primary-antibodies/ythdf3-antibody-hl2235-ab317173'>ab317173</a>) at 1/2000 dilution

Lane 1:

Wild-type PC-3 cell lysates at 20 µg

Lane 2:

Western blot - Human YTHDF3 knockout PC-3 cell line (<a href='/products/cell-lines/human-ythdf3-knockout-pc-3-cell-line-ab290587'>ab290587</a>) at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

HT-1080 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 64 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-YTHDF3 antibody [EPR21912-3] ab220161 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 70 kDa in Wild-type PC-3 cell lysates with no signal observed at this size in YTHDF3 knockout PC-3 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-YTHDF3 antibody [EPR21912-3] (<a href='/products/primary-antibodies/ythdf3-antibody-epr21912-3-ab220161'>ab220161</a>) at 1/1000 dilution

Lane 1:

Wild-type PC-3 cell lysates at 20 µg

Lane 2:

Western blot - Human YTHDF3 knockout PC-3 cell line (<a href='/products/cell-lines/human-ythdf3-knockout-pc-3-cell-line-ab290587'>ab290587</a>) at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

HT-1080 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 64 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

False colour image of Western blot : Anti-p53 antibody [Y5] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32049 was shown to bind specifically to p53. A band was observed at 50 kDa in wild-type HAP1 cell lysate with no signal observed at this size in tp53 knockout cell line. To generate this image, wild-type and tp53 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

This antibody recognises both wild-type and mutant forms of human p53 and is not specific to any particular mutation. Expression levels of p53 can vary significantly between cell lines, with mutant forms often showing higher stability and abundance. In low-expressing wild-type cell lines, p53 levels may be increased using treatments such as camptothecin or irinotecan.

All lanes:

Western blot - Anti-p53 antibody [Y5] (<a href='/products/primary-antibodies/p53-antibody-y5-ab32049'>ab32049</a>) at 1/1000 dilution

Lane 1:

Saos-2 cell lysate at 20 µg

Lane 2:

A431 cell lysate at 20 µg

Lane 3:

Wild-type HAP1 cell lysate at 20 µg

Lane 4:

TP53 knockout HAP1 cell lysate at 20 µg

Lane 5:

MCF7 cell lysate at 20 µg

Lane 6:

HEK-293T cell lysate at 20 µg

Secondary

Lanes 1 - 6:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 6:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 43 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-Glutaminase antibody [EP7212] ab156876 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. Three specific bands are detected by ab156876. We believe these correspond to isoform 1 (KGA), isoform 3 (GAC) and the mature mitochondrial subunits. Bands were observed at 68, 60 and 55 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in GLS knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Glutaminase antibody [EP7212] (<a href='/products/primary-antibodies/glutaminase-antibody-ep7212-ab156876'>ab156876</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 cell lysates at 20 µg

Lane 2:

Western blot - Human GLS knockout MCF7 cell line (<a href='/products/cell-lines/human-gls-knockout-mcf7-cell-line-ab287444'>ab287444</a>) at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

OVCAR-3 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 68 kDa,60 kDa,55 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-HEXB antibody [EPR7978] ab140649 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. Band were observed at 63 and 29 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in HEXB knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-HEXB antibody [EPR7978] (<a href='/products/primary-antibodies/hexb-antibody-epr7978-ab140649'>ab140649</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG cell lysates at 20 µg

Lane 2:

HEXB knockout U-87 MG cell lysates at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

Jurkat cell lysates at 20 µg

Lane 5:

Daudi cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 63 kDa

Observed band size: 63 kDa,29 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Rabbit monoclonal [EPR23696-22] to BMAL1 ab230822 staining at 1/500 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 69 kDa in Wild-type A549 and HeLa cell lysates with no signal observed at this size in ARNTL knockout A549 cell line and ARNTL knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-BMAL1 antibody [EPR23696-22] (<a href='/products/primary-antibodies/bmal1-antibody-epr23696-22-ab230822'>ab230822</a>) at 1/500 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

Western blot - Human ARNTL knockout A549 cell line (ab288835) at 20 µg

Lane 3:

Wild-type HeLa ab255928 cell lysates at 20 µg

Lane 4:

Western blot - Human ARNTL (BMAL1) knockout HeLa cell line (<a href='/products/cell-lines/human-arntl-bmal1-knockout-hela-cell-line-ab264701'>ab264701</a>) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 69 kDa

Observed band size: 69 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-PTEN antibody [Y184] ab32199 staining at 1/500 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 47 kDa in Wild-type A549 cell lysates with no signal observed at this size in PTEN knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PTEN antibody [Y184] (<a href='/products/primary-antibodies/pten-antibody-y184-ab32199'>ab32199</a>) at 1/500 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

Western blot - Human PTEN knockout A549 cell line (<a href='/products/cell-lines/human-pten-knockout-a549-cell-line-ab286704'>ab286704</a>) at 20 µg

Lane 3:

MCF7 cell lysates at 20 µg

Lane 4:

LNCaP cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-c-Rel antibody [EPR2559(2)] ab133251 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 75 kDa in Wild-type MCF7 nuclear fraction lysates with no signal observed at this size in REL knockout MCF7 nuclear fraction. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-c-Rel antibody [EPR2559(2)] (<a href='/products/primary-antibodies/c-rel-antibody-epr25592-ab133251'>ab133251</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7, nuclear fraction cell lysates at 20 µg

Lane 2:

Western blot - Human REL knockout MCF7 cell line (ab286278) at 20 µg

Lane 3:

Raji cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 69 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-PAI1 antibody [EPR17796] ab187263 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 45 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in SERPINE1 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PAI1 antibody [EPR17796] (<a href='/products/primary-antibodies/pai1-antibody-epr17796-ab187263'>ab187263</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG cell lysates at 20 µg

Lane 2:

Western blot - Human SERPINE1 knockout U-87 MG cell line (<a href='/products/cell-lines/human-serpine1-knockout-u-87-mg-cell-line-ab306851'>ab306851</a>) at 20 µg

Lane 3:

A549 cell lysates at 20 µg

Lane 4:

HEK-293 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-AP4E1 antibody [EPR28049-87] ab318189 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 127 kDa in Wild-type SH-SY5Y cell lysates with no signal observed at this size in AP4E1 knockout SH-SY5Y cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-AP4E1 antibody [EPR28049-87] (<a href='/products/primary-antibodies/ap4e1-antibody-epr28049-87-ab318189'>ab318189</a>) at 1/1000 dilution

Lane 1:

Wild-type SH-SY5Y cell lysates at 20 µg

Lane 2:

AP4E1 knockout SH-SY5Y cell lysates at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

PC-3 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 127 kDa

Observed band size: 127 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-PTEN antibody [EPR4408-76] ab133532 staining at 1/10000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 50 kDa in Wild-type A549 cell lysates with no signal observed at this size in PTEN knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PTEN antibody [EPR4408-76] (<a href='/products/primary-antibodies/pten-antibody-epr4408-76-ab133532'>ab133532</a>) at 1/10000 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

Western blot - Human PTEN knockout A549 cell line (<a href='/products/cell-lines/human-pten-knockout-a549-cell-line-ab286704'>ab286704</a>) at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Rabbit Monoclonal [EPR27024-81] to USP30 ab314749 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 59 kDa in Wild-type RAW 264.7 cell lysates with no signal observed at this size in USP30 knockout RAW 264.7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-USP30 antibody [EPR27024-81] (<a href='/products/primary-antibodies/usp30-antibody-epr27024-81-ab314749'>ab314749</a>) at 1/1000 dilution

Lane 1:

Wild-type RAW 264.7 cell lysates at 20 µg

Lane 2:

USP30 knockout RAW 264.7 cell lysates at 20 µg

Lane 3:

Wild-type A549 cell lysates at 20 µg

Lane 4:

USP30 knockout A549 <a href='/products/cell-lines/human-usp30-knockout-a549-cell-line-ab324260'>ab324260</a> cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 59 kDa

Observed band size: 59 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-Rab11A antibody [EPR28202-94] ab316151 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 24 kDa in Wild-type A549 cell lysates with no signal observed at this size in RAB11A knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Rab11A antibody [EPR28202-94] (<a href='/products/primary-antibodies/rab11a-antibody-epr28202-94-ab316151'>ab316151</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

RAB11A knockout A549 cell lysates at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

HCT 116 cell lysates at 20 µg

Lane 5:

LNCaP cell lysates at 20 µg

Lane 6:

Jurkat cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-PAI1 antibody [EPR21850-262] ab317604 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 45 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in SERPINE1 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PAI1 antibody [EPR21850-262] (<a href='/products/primary-antibodies/pai1-antibody-epr21850-262-ab317604'>ab317604</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG cell lysates at 20 µg

Lane 2:

Western blot - Human SERPINE1 knockout U-87 MG cell line (<a href='/products/cell-lines/human-serpine1-knockout-u-87-mg-cell-line-ab306851'>ab306851</a>) at 20 µg

Lane 3:

A549 cell lysates at 20 µg

Lane 4:

HEK-293 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-PAI1 antibody [EPR17795] ab187262 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 45 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in SERPINE1 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PAI1 antibody [EPR17795] (<a href='/products/primary-antibodies/pai1-antibody-epr17795-ab187262'>ab187262</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG cell lysates at 20 µg

Lane 2:

Western blot - Human SERPINE1 knockout U-87 MG cell line (<a href='/products/cell-lines/human-serpine1-knockout-u-87-mg-cell-line-ab306851'>ab306851</a>) at 20 µg

Lane 3:

A549 cell lysates at 20 µg

Lane 4:

HEK-293 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-AKT1 antibody [HL1142] ab308343 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 56 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in AKT1 knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-AKT1 antibody [HL1142] (<a href='/products/primary-antibodies/akt1-antibody-hl1142-ab308343'>ab308343</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysates at 20 µg

Lane 2:

Western blot - Human AKT1 knockout HCT116 cell line (<a href='/products/cell-lines/human-akt1-knockout-hct116-cell-line-ab286570'>ab286570</a>) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 56 kDa

Observed band size: 56 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Rabbit monoclonal [EPR4204] to LAMP1 ab108597 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 80-120 kDa in Wild-type A549 and HCT 116 cell lysates with no signal observed at this size in LAMP1 knockout A549 cell line and LAMP1 knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-LAMP1 antibody [EPR4204] - Lysosome Marker (<a href='/products/primary-antibodies/lamp1-antibody-epr4204-lysosome-marker-ab108597'>ab108597</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

Western blot - Human LAMP1 knockout A549 cell line (<a href='/products/cell-lines/human-lamp1-knockout-a549-cell-line-ab288919'>ab288919</a>) at 20 µg

Lane 3:

Wild-type HCT 116 ab277289 cell lysates at 20 µg

Lane 4:

Western blot - Human LAMP1 knockout HCT116 cell line (<a href='/products/cell-lines/human-lamp1-knockout-hct116-cell-line-ab289151'>ab289151</a>) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 80-120 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-ER81/ETV1 antibody [EPR28393-132] ab314874 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 60 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in ETV1 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-ER81/ETV1 antibody [EPR28393-132] (<a href='/products/primary-antibodies/er81-etv1-antibody-epr28393-132-ab314874'>ab314874</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG cell lysates at 20 µg

Lane 2:

ETV1 knockout U-87 MG cell lysates at 20 µg

Lane 3:

SH-SY5Y cell lysates at 20 µg

Lane 4:

MCF7 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 60 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-ADAM10 antibody [EPR5622] ab124695 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 84 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in ADAM10 knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-ADAM10 antibody [EPR5622] (<a href='/products/primary-antibodies/adam10-antibody-epr5622-ab124695'>ab124695</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 cell lysates at 20 µg

Lane 2:

Western blot - Human ADAM10 knockout MCF7 cell line (ab287197) at 20 µg

Lane 3:

Jurkat cell lysates at 20 µg

Lane 4:

U-2 OS cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 84 kDa

Observed band size: 84 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-LAMP1 antibody [RM1217] - Lysosome Marker ab320851 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 80-120 kDa in Wild-type A549 and HCT 116 cell lysates with no signal observed at this size in LAMP1 knockout A549 cell line and LAMP1 knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-LAMP1 antibody [RM1217] - Lysosome Marker (<a href='/products/primary-antibodies/lamp1-antibody-rm1217-lysosome-marker-ab320851'>ab320851</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

Western blot - Human LAMP1 knockout A549 cell line (<a href='/products/cell-lines/human-lamp1-knockout-a549-cell-line-ab288919'>ab288919</a>) at 20 µg

Lane 3:

Wild-type HCT 116 ab277289 cell lysates at 20 µg

Lane 4:

Western blot - Human LAMP1 knockout HCT116 cell line (<a href='/products/cell-lines/human-lamp1-knockout-hct116-cell-line-ab289151'>ab289151</a>) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 80-120 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-PTEN antibody [SP218] - C-terminal ab228466 staining at 1/400 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 47 kDa in Wild-type HeLa cell lysates with no signal observed at this size in PTEN knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PTEN antibody [SP218] - C-terminal (<a href='/products/primary-antibodies/pten-antibody-sp218-c-terminal-ab228466'>ab228466</a>) at 1/400 dilution

Lane 1:

Wild-type HeLa cell lysates at 20 µg

Lane 2:

Western blot - Human PTEN knockout HeLa cell line (<a href='/products/cell-lines/human-pten-knockout-hela-cell-line-ab255419'>ab255419</a>) at 20 µg

Lane 3:

MCF7 cell lysates at 20 µg

Lane 4:

LNCaP cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-FACL4 antibody [EPR17587] ab205197 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 79 kDa in Wild-type A431 cell lysates with no signal observed at this size in ACSL4 knockout A431 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-FACL4 antibody [EPR17587] (<a href='/products/primary-antibodies/facl4-antibody-epr17587-ab205197'>ab205197</a>) at 1/1000 dilution

Lane 1:

Wild-type A431 cell lysates at 20 µg

Lane 2:

Western blot - Human ACSL4 (FACL4) knockout A-431 cell line (<a href='/products/cell-lines/human-acsl4-facl4-knockout-a-431-cell-line-ab269613'>ab269613</a>) at 20 µg

Lane 3:

HEK-293 cell lysates at 20 µg

Lane 4:

K562 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 79 kDa

Observed band size: 79 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Rabbit monoclonal [EPR24395-31] to LAMP1 ab278043 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 80-120 kDa in Wild-type A549 and HCT 116 cell lysates with no signal observed at this size in LAMP1 knockout A549 cell line and LAMP1 knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

Lanes 1 - 4:

Western blot - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (<a href='/products/primary-antibodies/lamp1-antibody-epr24395-31-bsa-and-azide-free-ab278046'>ab278046</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (<a href='/products/primary-antibodies/lamp1-antibody-epr24395-31-lysosome-marker-ab278043'>ab278043</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

Western blot - Human LAMP1 knockout A549 cell line (<a href='/products/cell-lines/human-lamp1-knockout-a549-cell-line-ab288919'>ab288919</a>) at 20 µg

Lane 3:

Wild-type HCT 116 ab277289 cell lysates at 20 µg

Lane 4:

Western blot - Human LAMP1 knockout HCT116 cell line (<a href='/products/cell-lines/human-lamp1-knockout-hct116-cell-line-ab289151'>ab289151</a>) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 80-120 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-Ferritin Light Chain antibody [CPT-R10-58-11] ab243096 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 19 kDa in Wild-type HeLa cell lysates with no signal observed at this size in FTL knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Ferritin Light Chain antibody [CPT-R10-58-11] (<a href='/products/primary-antibodies/ferritin-light-chain-antibody-cpt-r10-58-11-ab243096'>ab243096</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysates at 20 µg

Lane 2:

Western blot - Human FTL knockout HeLa cell line (<a href='/products/cell-lines/human-ftl-knockout-hela-cell-line-ab265533'>ab265533</a>) at 20 µg

Lane 3:

HepG2 cell lysates at 20 µg

Lane 4:

T-47D cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 20 kDa

Observed band size: 19 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-DDR2 antibody [EPR18826-119] ab280354 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 120 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-DDR2 antibody [EPR18826-119] (<a href='/products/primary-antibodies/ddr2-antibody-epr18826-119-ab280354'>ab280354</a>) at 1/1000 dilution

Lane 1:

SH-SY5Y not boiled cell lysates at 20 µg

Lane 2:

U-2 OS not boiled cell lysates at 20 µg

Lane 3:

Calu-3 not boiled cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 97 kDa

Observed band size: 120 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-MFI2/MTf antibody [EPR27232-23] ab303514 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 80 kDa in Wild-type A549 cell lysates with no signal observed at this size in MELTF knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-MFI2/MTf antibody [EPR27232-23] (<a href='/products/primary-antibodies/mfi2-mtf-antibody-epr27232-23-ab303514'>ab303514</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 not boiled cell lysates at 20 µg

Lane 2:

MELTF knockout A549 not boiled cell lysates at 20 µg

Lane 3:

A431 not boiled cell lysates at 20 µg

Lane 4:

LNCaP not boiled cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 80 kDa

Observed band size: 80 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-ER81/ETV1 antibody [DZR-1-29] ab322139 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 60 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in ETV1 knockout U-87 MG cell line. The band observed in the CRISPR-Cas9 edited lysate lane below 60 kDa is likely to represent a truncated form of ETV1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-ER81/ETV1 antibody [DZR-1-29] (<a href='/products/primary-antibodies/er81-etv1-antibody-dzr-1-29-ab322139'>ab322139</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG cell lysates at 20 µg

Lane 2:

ETV1 CRISPR-Cas9 edited U-87 MG cell lysates at 20 µg

Lane 3:

SH-SY5Y cell lysates at 20 µg

Lane 4:

MCF7 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 60 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-Glutaminase C antibody [EPR19525] ab202027 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 60 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in GLS knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Glutaminase C antibody [EPR19525] (<a href='/products/primary-antibodies/glutaminase-c-antibody-epr19525-ab202027'>ab202027</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 cell lysates at 20 µg

Lane 2:

Western blot - Human GLS knockout MCF7 cell line (<a href='/products/cell-lines/human-gls-knockout-mcf7-cell-line-ab287444'>ab287444</a>) at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

OVCAR-3 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 60 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-PTEN antibody [EPR9941] ab154812 staining at 1/10000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 50 kDa in Wild-type A549 and HeLa cell lysates with no signal observed at this size in PTEN knockout A549 cell line and PTEN knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PTEN antibody [EPR9941] (<a href='/products/primary-antibodies/pten-antibody-epr9941-ab154812'>ab154812</a>) at 1/10000 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

Western blot - Human PTEN knockout A549 cell line (<a href='/products/cell-lines/human-pten-knockout-a549-cell-line-ab286704'>ab286704</a>) at 20 µg

Lane 3:

Wild-type HeLa ab255928 cell lysates at 20 µg

Lane 4:

Western blot - Human PTEN knockout HeLa cell line (<a href='/products/cell-lines/human-pten-knockout-hela-cell-line-ab255419'>ab255419</a>) at 20 µg

Lane 5:

MCF7 cell lysates at 20 µg

Lane 6:

LNCaP cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade ab1791 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 18 kDa in Wild-type MCF7 cell lysates with less signal observed at this size in H3C1 knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (<a href='/products/primary-antibodies/histone-h3-antibody-nuclear-marker-and-chip-grade-ab1791'>ab1791</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 cell lysates at 40 µg

Lane 2:

H3C1 knockout MCF7 cell lysates at 40 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

A549 Membrane cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 15 kDa

Observed band size: 18 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Lanes 1- 2 : Merged signal (red and green). Green - ab109440 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab109440 was shown to react with NFkB p100/NFKB2 in wild-type HepG2 cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab262323 (CRISPR/Cas9 edited cell lysate ab257247) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HepG2 and NFKB2 CRISPR/Cas9 edited HepG2 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109440 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (<a href='/products/primary-antibodies/nfkb-p100-nfkb2-antibody-epr4686-ab109440'>ab109440</a>) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (<a href='/products/cell-lysates/human-nfkb2-nfkb-p100-nfkb2-knockout-hep-g2-cell-lysate-ab257247'>ab257247</a>) at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 30 kDa,97 kDa

Observed band size: 120 kDa,30 kDa,37 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-KMT6 / EZH2 antibody [SP129] ab227648 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 85 kDa in Wild-type A375 cell lysates with no signal observed at this size in EZH2 knockout A375 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-KMT6 / EZH2 antibody [SP129] (<a href='/products/primary-antibodies/kmt6-ezh2-antibody-sp129-ab227648'>ab227648</a>) at 1/1000 dilution

Lane 1:

Wild-type A375 cell lysates at 20 µg

Lane 2:

EZH2 knockout A375 cell lysates at 20 µg

Lane 3:

HEK-293 cell lysates at 20 µg

Lane 4:

MCF7 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 85 kDa

Observed band size: 85 kDa

false