Anti-GAPDH 抗体 [EPR16891] - Loading Control

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocyte of rat spleen is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling GAPDH with ab181602 at 1/180 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Immunocytochemistry/immunofluorescence staining of 4% paraformaldehyde fixed; 0.1% triton X 100 permeabilized HeLa (human cervix adenocarcinoma) cells labeling GAPDH with ab181602 at dilution of 1/500. The secondary antibody used was Alexa Fluor^® 488; goat anti-rabbit IgG (ab150077) at a dilution of 1/400. Nucleus was counter-stained with DAPI (blue). ab7291, a mouse anti-tubulin antibody (1/500) was used to stain tubulin along with ab150120 (AlexaFluor^®594 goat anti-mouse secondary, 1/500). The negative controls are shown in the bottom middle and right hand panels- for negative control 1 primary antibody (ab181602; 1/500) and secondary antibody (ab150120; 1/500) was used. For negative control 2 primary antibody (ab7291; 1/500) and secondary antibody (ab150077; 1/400) was used.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic and nucleus staining on the tumor cells of transitional cell carcinoma of human bladder is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

STT3A was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) whole cell lysate with ab320831 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab320831 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Immunoprecipitation, ab320831 was shown to bind specifically to STT3A. Target of interest was enriched at 60 kDa in wild-type HEK-293T cell lysates (lane 2) with no signal observed at this size in STT3A knockout cell line (lane 5) (lane 5, knockout cell line ab266320 / knockout cell lysate ab259164).
The bands above 60kDa are likely to be aggregation, and are absent in the KO samples. Unlike the Western Blot data, these samples have been boiled. ab181602 was used as a GAPDH loading control.

All lanes:

Immunoprecipitation - Anti-STT3A antibody [EPR29178-10] (<a href='/products/primary-antibodies/stt3a-antibody-epr29178-10-ab320831'>ab320831</a>) at 1/1000 dilution

Lane 1:

Parental HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 10 µg

Lane 2:

Parental HEK-293T (human embryonic kidney epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/products/primary-antibodies/stt3a-antibody-epr29178-10-ab320831'>ab320831</a> in Parental HEK-293T whole cell lysate

Lane 4:

STT3A KO HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 10 µg

Lane 5:

Immunoprecipitation - Human STT3A knockout HEK-293T cell line (<a href='/products/cell-lines/human-stt3a-knockout-hek-293t-cell-line-ab266320'>ab266320</a>)

Lane 6:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/products/primary-antibodies/stt3a-antibody-epr29178-10-ab320831'>ab320831</a> in STT3A KO HEK-293T whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 60 kDa

false

Exposure time: 84s

• IP

Supplier Data

Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Dnmt3a was immunoprecipitated from 0.35 mg Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab323708 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323708 at 1000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Dnmt3a antibody [EPR29184-89] (<a href='/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</a>) at 1/1000 dilution

Lane 1:

(Input) Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

<a href='/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</a> at 1/30 IP in Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</a> in parental HeLa whole cell lysate at 10 µg

Lane 4:

(Input) DNMT3A knockout HeLa whole cell lysate at 10 µg

Lane 5:

<a href='/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</a> at 1/30 IP in DNMT3A knockout HeLa whole cell lysate at 10 µg

Lane 6:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/products/primary-antibodies/dnmt3a-antibody-epr29184-89-ab323708'>ab323708</a> in DNMT3A knockout HeLa whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 130 kDa,36 kDa

false

Exposure time: 24s

• IP

Supplier Data

Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

GAPDH was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181602 at 1/60 dilution. Western blot was performed from the immunoprecipitate using ab181602 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1 : HeLa whole cell extract.

Lane 2 : PBS instead of HeLa whole cell extract.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

Predicted band size: 36 kDa

Observed band size: 36 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the bands higher than 45 kDa is unknown.

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-TMEM175 antibody [RM1351] (<a href='/products/primary-antibodies/tmem175-antibody-rm1351-ab325021'>ab325021</a>) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

MEF (mouse embryo fibroblast) whole cell lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 4:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 5:

Human lung tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 45 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : WI-38

Samples are non-boiled as boiling may cause protein aggregation.

Lanes 2-3 developed were using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

Lane1 was incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and lanes 2-3 were incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-SorCS1 antibody [EPR30536-522] (ab325598) at 1/1000 dilution

Lane 1:

Human hypothalamus tissue lysate at 20 µg

Lane 2:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

WI-38 (human fetal lung fibroblast) whole cell lysate at 40 µg

Secondary

Lane 1:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 2 - 3:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 130 kDa,36 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : HepG2

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-4 : 37 seconds, lanes 5-6 : 15 seconds

All lanes:

Western blot - Anti-EDG2/LPA-1 antibody [EPR30753-555] (<a href='/products/primary-antibodies/edg2-lpa-1-antibody-epr30753-555-ab325552'>ab325552</a>) at 1/1000 dilution

Lane 1:

U-118 MG (human brain glioblastoma cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

U-118 MG transfected with siRNA specifically targeting LPAR1 whole cell lysate at 20 µg

Lane 3:

Untreated U-118 MG whole cell lysate at 20 µg

Lane 4:

U-118 MG whole cell lysate treated with Protein Deglycosylation Peptide:N-glycosidase F (PNGase F) at 20 µg

Lane 5:

MG-63 (human osteosarcoma fibroblast) whole cell lysate at 20 µg

Lane 6:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 25-75 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human TMEM106C and human TMEM106A by western blot.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);Anti-Myc tag® antibody (1 : 5000) (40KDa);.

All lanes:

Western blot - Anti-TMEM106B antibody [EPR30569-44] - C-terminal (ab325607) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 2:

293T transfected with a human TMEM106B expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with a human TMEM106C expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 4:

293T transfected with a human TMEM106A expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 40 kDa,36 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

ab181602 was used as a GAPDH loading control at a 1/200000 dilution.

Exposure time :
Lane 1-2 : 180 seconds;
Lane 3-4 : 26 seconds

All lanes:

Western blot - Anti-PAI1 antibody [EPR21850-262] - BSA and Azide free (Detector) (<a href='/products/primary-antibodies/pai1-antibody-epr21850-262-bsa-and-azide-free-detector-ab269409'>ab269409</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Mouse placenta tissue lysate at 20 µg

Lane 4:

Rat placenta tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

In Western blot, ab325523 was shown to bind specifically to Mineralocorticoid Receptor. Target of interest was observed at 107 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in Mineralocorticoid Receptor knockout cell line (lane 2, knockout cell line ab265540 / knockout cell lysate ab257279).

To minimize protein degradation, tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the lower MW bands below 107 kDa are unknown.

All lanes:

Western blot - Anti-Mineralocorticoid Receptor antibody [EPR29519-576] (<a href='/products/primary-antibodies/mineralocorticoid-receptor-antibody-epr29519-576-ab325523'>ab325523</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 48 µg

Lane 2:

Mineralocorticoid Receptor knockout HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 107 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human IMPDH2 by western blot.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-IMPDH1 antibody [EPR30383-519] (<a href='/products/primary-antibodies/impdh1-antibody-epr30383-519-ab325431'>ab325431</a>) at 1/1000 dilution

Lane 1:

His-tag purified protein overexpressed in 293T human embryonic kidney epithelial cells, transfected with a human IMPDH1 expression vector incorporating a Myc-His® tag, whole-cell lysate at 20 µg

Lane 2:

His-tag purified protein overexpressed in 293T human embryonic kidney epithelial cells, transfected with a human IMPDH2 expression vector incorporating a Myc-His® tag, whole-cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 55 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : CTLL-2 (PMID : 1396582). In Western blot, anti-GAPDH antibody (ab181602) staining at 1/20, 0000 dilution. Exposure time : 125 seconds

All lanes:

Western blot - Anti-PD1 antibody [RM1033] (<a href='/products/primary-antibodies/pd1-antibody-rm1033-ab309363'>ab309363</a>) at 1/1000 dilution

Lane 1:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 2:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 3:

EL4.IL-2 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 4:

CTLL-2 (mouse T lymphocyte) whole cell lysate at 20 µg

Lane 5:

Human tomsil tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60 kDa,80 kDa

false

Exposure time: 125s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as GAPDH loading control. ab176842 was for Histone H3 detection.

All lanes:

Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (<a href='/products/primary-antibodies/histone-h3-acetyl-k9-antibody-y28-chip-grade-ab32129'>ab32129</a>) at 1/10000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, ab309157 was shown to bind specifically to Alpha-synuclein.  Target of interest was observed at 18 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in Alpha-synuclein knockout cell line (lane 2). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds.

All lanes:

Western blot - Anti-Alpha-synuclein antibody [MJFR1] - Mouse IgG2a (Chimeric) (<a href='/products/primary-antibodies/alpha-synuclein-antibody-mjfr1-mouse-igg2a-chimeric-ab309157'>ab309157</a>) at 1/1000 dilution

Lane 1:

Parental HAP1 whole cell lysate at 20 µg

Lane 2:

Alpha-synuclein knockout HAP1 whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Observed band size: 18 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with recombinant human SORCS3 by western blot.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (130KDa).

All lanes:

Western blot - Anti-SorCS1 antibody [EPR30536-522] (ab325598) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T transfected with a human SORCS1 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T transfected with a human SORCS3 expression vector containing a His-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 130 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

In Western blot, ab184557 was shown to bind specifically to SMAD2. Target of interest was observed at 58 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in SMAD2 knockout cell line (lane 2) (lane 2, knockout cell line ab255430).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Smad2 (phospho S250) antibody [EPR2855(2)] (<a href='/products/primary-antibodies/smad2-phospho-s250-antibody-epr28552-ab184557'>ab184557</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SMAD2 knockout HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 58 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Lamin B1 antibody [EPR22165-121] (ab229025) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Histone H3 antibody [RM1288] - Nuclear Marker (<a href='/products/primary-antibodies/histone-h3-antibody-rm1288-nuclear-marker-ab322707'>ab322707</a>) at 1/10000 dilution

Lane 1:

HEK-293 (human embryonic kidney epithelial cell) nuclear fraction at 10 µg

Lane 2:

HEK-293 cytoplasmic fraction at 10 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) nuclear fraction at 10 µg

Lane 4:

NIH/3T3 cytoplasmic fraction at 10 µg

Lane 5:

Rat brain tissue nuclear fraction at 10 µg

Lane 6:

Rat brain tissue cytoplasmic fraction at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 16 kDa,14 kDa,36 kDa,66 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602).

Negative control : liver.

All lanes:

Western blot - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (<a href='/products/primary-antibodies/gfap-antibody-epr1034y-astrocyte-marker-chicken-igy-chimeric-ab323239'>ab323239</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat liver tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/10000 dilution

Observed band size: 45-50 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Exposure time : Lane 1 : 10 seconds; Lane 2 : 180 seconds

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining.

Negative control : HepG2 (PMID : 28378740).

Samples are non-boiled as boiling may cause protein aggregates.

All lanes:

Western blot - Anti-CD47 antibody [EPR24922-21] (<a href='/products/primary-antibodies/cd47-antibody-epr24922-21-ab300124'>ab300124</a>) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

U937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 4:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

Lanes 1 - 5:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lanes 1 - 5:

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/100000 dilution

Observed band size: 47-52 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : skeletal muscle.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PHGDH/Malate dehydrogenase antibody [EPR28634-1] (<a href='/products/primary-antibodies/phgdh-malate-dehydrogenase-antibody-epr28634-1-ab324530'>ab324530</a>) at 1/1000 dilution

Lane 1:

Mouse cerebellum tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle lysat at 20 µg

Lane 3:

Human cerebellum tissue lysate at 20 µg

Lane 4:

Human skeletal muscle lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 57 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (<a href='/products/primary-antibodies/adenine-nucleotide-translocator-2-ant-2-antibody-epr29813-534-ab325120'>ab325120</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting SLC25A5 whole cell lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 6:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 32 kDa,36 kDa

false

Exposure time: 125s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : Daudi (PMID : 35316216).

Low expression : A549 (PMID : 30250195).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PHGDH/Malate dehydrogenase antibody [EPR28634-1] (<a href='/products/primary-antibodies/phgdh-malate-dehydrogenase-antibody-epr28634-1-ab324530'>ab324530</a>) at 1/1000 dilution

Lane 1:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 3:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 6:

Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 7:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 8:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 9:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 57 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (<a href='/products/primary-antibodies/adenine-nucleotide-translocator-2-ant-2-antibody-epr29813-534-ab325120'>ab325120</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole lysate at 20 µg

Lane 2:

HeLa non-mitochondrial fraction at 20 µg

Lane 3:

HeLa mitochondrial fraction at 20 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 32 kDa,36 kDa,16 kDa

false

Exposure time: 125s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution (36KDa).

In Western blot, Anti-6X His tag® antibody [AD1.1.10] total protein control (1/1000) (30KDa)

All lanes:

Western blot - Anti-Langerin antibody [EPR24685-12] (<a href='/products/primary-antibodies/langerin-antibody-epr24685-12-ab283686'>ab283686</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T transfected with a human CD207/Langerin expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T transfected with a mouse CD207/Langerin expression vector containing a His-tag, whole cell lysate (Low Expression) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 40 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 12423348; PMID : 18256700; PMID : 9733783).

Cystatin F is a glycoprotein of approximately 18-20 kDa and detected as 15 kDa band after treated with Deglycosylation Peptide : N-glycosidase F (PNGase F).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Cystatin F antibody [EPR29572-528] (<a href='/products/primary-antibodies/cystatin-f-antibody-epr29572-528-ab324347'>ab324347</a>) at 1/1000 dilution

Lane 1:

Untreated U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 2:

U-937 whole cell lysate treated with Protein Deglycosylation Peptide:N-glycosidase F (PNGase F) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 18-20 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : HeLa, 293T (PMID : 29180998); K-562 (PMID : 33291222); THP-1, MOLT-4 (PMID : 9733783)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 12423348; PMID : 18256700; PMID : 9733783; PMID : 33291222).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-2 : 180 seconds; Lanes 3-8 : 15 seconds

All lanes:

Western blot - Anti-Cystatin F antibody [EPR29572-528] (<a href='/products/primary-antibodies/cystatin-f-antibody-epr29572-528-ab324347'>ab324347</a>) at 1/1000 dilution

Lane 1:

No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) whole cell lysate at 20 µg

Lane 2:

HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg

Lane 3:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 4:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 6:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 7:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 8:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 18-20 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-MR1 antibody [EPR30434-652] (ab325648) at 1/1000 dilution

Lane 1:

T-47D (human ductal breast epithelial tumor epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

T-47D transfected with siRNA specifically targeting MR1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 26 kDa,38 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

U-118MG cells carry the FIG-ROS1 fusion (PMID : 12661006). Tyrosine phosphorylated ROS1 is downregulated in response to crizotinib treatment (PMID : 24971615)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Phospho-ROS1 (Tyr2274/2334) antibody [EPR28163-133] (<a href='/products/primary-antibodies/phospho-ros1-tyr2274-2334-antibody-epr28163-133-ab324617'>ab324617</a>) at 1/1000 dilution

Lane 1:

Untreated U-118 MG (human brain glioblastoma cell) whole cell lysate (untreated membrane) at 50 µg

Lane 2:

U-118 MG treated with 250 nM crizotinib for 4 hours whole cell lysate (untreated membrane) at 50 µg

Lane 3:

Untreated U-118 MG whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 4:

U-118 MG treated with 250 nM crizotinib for 4 hours whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 110 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : TT.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

The identity of the bands higher 30 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-UCHL3 antibody [EPR29841-548] (<a href='/products/primary-antibodies/uchl3-antibody-epr29841-548-ab323384'>ab323384</a>) at 1/1000 dilution

Lane 1:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 50 µg

Lane 3:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 50 µg

Lane 4:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 5:

TT (human thyroid carcinoma epithelial cell) whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 26 kDa,36 kDa

true

Exposure time: 92s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and dilution buffer and concentration : 5% NFDM/TBST

Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) was used as a loading control at 1/200000 dilution.

Low expression : HeLa.
Negative control : HepG2.

All lanes:

Western blot - Anti-HLA DR + DP + DQ antibody [CR3/43] (<a href='/products/primary-antibodies/hla-dr-dp-dq-antibody-cr3-43-ab7856'>ab7856</a>) at 1/1000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

KARPAS-299 (human T cell lymphoma cell) whole cell lysate at 20 µg

Lane 3:

HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate at 20 µg

Lane 4:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 30 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression of Thymosin beta 4 is upregulated in response to PMA treatment. (PMID : 3500230).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Thymosin beta 4 antibody [EPR26609-45] (<a href='/products/primary-antibodies/thymosin-beta-4-antibody-epr26609-45-ab323478'>ab323478</a>) at 1/1000 dilution

Lane 1:

Untreated HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 40 µg

Lane 2:

HL-60 treated with 50ng/ml PMA for 72h whole cell lysate at 40 µg

Lane 3:

HL-60 treated with 10 uM MG-132 for 4 hours whole cell lysate at 40 µg

Lane 4:

HL-60 treated with 50ng/ml PMA for 72h, 10 uM MG-132 was then added for additional 4h whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 13 kDa,36 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 1% BSA/TBST.

Performed under reducing conditions.

In Western blot, ab323846 was shown to bind specifically to Rb. Target of interest was observed at 106 kDa in Calyculin A treated wild-type HAP1 cell lysates (lane 2) with no signal observed at this size in Calyculin A treated Rb knockout cell line (lane 4).

Lanes 1-4 on the left untreated membrane with alkaline phosphatase; Lanes 1-4 on the right treated membrane with alkaline phosphatase.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Rb antibody [EPR17512] - (ab181616) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Rb (phospho T821 + T826) antibody [EPR28648-126] (<a href='/products/primary-antibodies/rb-phospho-t821-t826-antibody-epr28648-126-ab323846'>ab323846</a>) at 1/1000 dilution

Lane 1:

Untreated Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate (left: untreated membrane with alkaline phosphatase) at 20 µg

Lane 1:

Untreated Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate (right: treated membrane with alkaline phosphatase) at 20 µg

Lane 2:

Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (left: untreated membrane with alkaline phosphatase) at 20 µg

Lane 2:

Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (right: treated membrane with alkaline phosphatase) at 20 µg

Lane 3:

Untreated Rb knockout HAP1 whole cell lysate (left: untreated membrane with alkaline phosphatase) at 20 µg

Lane 3:

Untreated Rb knockout HAP1 whole cell lysate (right: treated membrane with alkaline phosphatase) at 20 µg

Lane 4:

Rb knockout HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (left: untreated membrane with alkaline phosphatase) at 20 µg

Lane 4:

Rb knockout HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (right: treated membrane with alkaline phosphatase) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 106 kDa,36 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and dilution buffer and concentration : 5% NFDM/TBST

Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) was used as a loading control at 1/200000 dilution.

Low expression : skeletal muscle.
The identity of MW band at approximately 50 kDa (in lane 2) is unknown.

All lanes:

Western blot - Anti-HLA DR + DP + DQ antibody [CR3/43] (<a href='/products/primary-antibodies/hla-dr-dp-dq-antibody-cr3-43-ab7856'>ab7856</a>) at 1/1000 dilution

Lane 1:

Human tonsil tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 30 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The observed molecular weight is greater than the theoretical value due to its hydrophilic nature, which causes slower migration on SDS-PAGE (PMID : 24109178).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Thymosin beta 4 antibody [EPR26609-45] (<a href='/products/primary-antibodies/thymosin-beta-4-antibody-epr26609-45-ab323478'>ab323478</a>) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

THP-1 transfected with siRNA specifically targeting Thymosin beta 4 whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 13 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab323371 was shown to bind specifically to TNFRSF10B. Target of interest was observed at 40, 48 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in TNFRSF10B knockout cell line (lane 2) (lane 2, knockout cell line ab264922).

The lanes 1-5 were developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

Doxorubicin treatment elevated the expression of DR5 (PMID : 12496481; PMID : 11468181; PMID : 11090076).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane1-4 : 103 seconds; lane 5-6 : 59 seconds

All lanes:

Western blot - Anti-TNFRSF10B antibody [RM2078] (<a href='/products/primary-antibodies/tnfrsf10b-antibody-rm2078-ab323371'>ab323371</a>) at 1/1000 dilution

Lane 1:

Parental HeLa (human cervical adenocarcinoma epithelial cell) whole lysate at 20 µg

Lane 2:

TNFRSF10B knockout HeLa whole lysate at 20 µg

Lane 3:

HT-1080 (human fibrosarcoma epithelial cell) whole lysate at 20 µg

Lane 4:

HepG2 (human hepatocelullar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Untreated HCT 116 (human colorectal carcinoma epithelial cell) whole lysate at 20 µg

Lane 6:

HCT 116 treated with 0.5uM doxorubicin for 24 hours whole lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 40 kDa,48 kDa,36 kDa

true

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Chk1 (phospho S345) antibody [EPR30270-566] (<a href='/products/primary-antibodies/chk1-phospho-s345-antibody-epr30270-566-ab325675'>ab325675</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T transfected with a human wild-type Chk1 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with a human Chk1 (S345A mutation) expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 56 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602)

Blocking and diluting buffer and concentration : QD20250827IMG15.

Low expression control : LNCaP(PMID : 15833880), 22Rv1(PMID : 16102578).

Performed under reducing conditions.

In Western blot, ab324683 was shown to bind specifically to JAK1. Target of interest was observed at 133 kDa in wild-type HAP1 cell lysates (lane 7) with no signal observed at this size in JAK1 knockout cell line (lane 8).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1-6 : 92 seconds Lane 7, 8 : 180 seconds

All lanes:

Western blot - Anti-JAK1 antibody [EPR24649-58] (<a href='/products/primary-antibodies/jak1-antibody-epr24649-58-ab324683'>ab324683</a>) at 1/1000 dilution

Lane 1:

HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate at 50 µg

Lane 2:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 3:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 4:

22Rv1 (human prostate carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 5:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 50 µg

Lane 6:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 7:

parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 50 µg

Lane 8:

JAK1 KO HAP1 whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 133 kDa,36 kDa

false