重组Anti-TNFAIP3抗体[EPR2663]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling TNFAIP3 with unpurified ab92324 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling TNFAIP3 with purified ab92324 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

• WB

Lab

Western blot - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

Western blot : Anti-TNFAIP3 antibody [EPR2663] (ab92324) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab92324 was shown to bind specifically to TNFAIP3. A band was observed at 90 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in TNFAIP3 knockout cell line. To generate this image, wild-type and TNFAIP3 knockout U-87 MG 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/1000 dilution

Lane 1:

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

Lane 2:

TNFAIP3 knockout U-87 MG cell lysate at 20 µg

Lane 3:

Wild-type A549 cell lysate at 20 µg

Lane 4:

TNFAIP3 knockout A549 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

false

• WB

Lab

Western blot - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

Lanes 1-4 : Merged signal (red and green). Green - ab92324 observed at 80 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab92324 Anti-TNFAIP3 antibody [EPR2663] was shown to specifically react with TNFAIP3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265983 (knockout cell lysate ab257112) was used. Wild-type and TNFAIP3 knockout samples were subjected to SDS-PAGE. ab92324 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TNFAIP3 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TNFAIP3 knockout HeLa cell line (<a href='/products/cell-lines/human-tnfaip3-knockout-hela-cell-line-ab265983'>ab265983</a>)

Lane 3:

Jurkat cell treated with 5ng/ml PMA for 48 hours and then treated with 2µg/ml PHA for 48 hours, whole cell lysate at 20 µg

Lane 4:

Untreated Jurkat cell lysate at 20 µg

Predicted band size: 89 kDa

Observed band size: 80 kDa

false

• WB

Lab

Western blot - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/3000 dilution

All lanes:

Jurkat cell lysate - treated with TNF (<a href='/products/proteins-peptides/recombinant-human-tnf-alpha-protein-ab9642'>ab9642</a>) and TPA at 10 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 89 kDa

Observed band size: 80 kDa

false

• WB

Supplier Data

Western blot - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

Lanes 1-4 : Merged signal (red and green). Green - ab92324 observed at 80 kDa. Red - loading control ab8245 observed at 36 kDa.

ab92324 Anti-TNFAIP3 antibody [EPR2663] was shown to specifically react with TNFAIP3 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266945 (knockout cell lysate ab257113) was used. Wild-type and TNFAIP3 knockout samples were subjected to SDS-PAGE. ab92324 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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-TNFAIP3 antibody [EPR2663] (ab92324) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

TNFAIP3 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TNFAIP3 knockout A549 cell line (<a href='/products/cell-lines/human-tnfaip3-knockout-a549-cell-line-ab266945'>ab266945</a>)

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) cell treated with 5ng/ml PMA for 48 hours and then treated with 2µg/ml PHA for 48 hours, whole cell lysate at 20 µg

Lane 4:

Untreated Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

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 1/10000 dilution

Predicted band size: 89 kDa

Observed band size: 80 kDa

false

• WB

Unknown

Western blot - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

All lanes:

Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/1000 dilution

Lane 1:

Jurkat cells treated with TNF (<a href='/products/proteins-peptides/recombinant-human-tnf-alpha-protein-ab9642'>ab9642</a>) and TPA at 10 µg

Lane 2:

Daudi cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 89 kDa

false

• WB

Lab

Western blot - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

Lanes 1- 4 : Merged signal (red and green). Green - ab92324 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab92324 was shown to react with TNFAIP3 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab266946 (knockout cell lysate ab257114) was used. Wild-type A549 and TNFAIP3 knockout A549 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. ab92324 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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-TNFAIP3 antibody [EPR2663] (ab92324) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

TNFAIP3 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human TNFAIP3 knockout A549 cell line (<a href='/products/cell-lines/human-tnfaip3-knockout-a549-cell-line-ab266946'>ab266946</a>)

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

TNFAIP3 knockout HeLa cell lysate at 20 µg

Predicted band size: 89 kDa

Observed band size: 90 kDa

false

• WB

Lab

Western blot - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/2000 dilution

All lanes:

Jurkat cell lysate - treated with TNF (<a href='/products/proteins-peptides/recombinant-human-tnf-alpha-protein-ab9642'>ab9642</a>) and TPA at 10 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 89 kDa

Observed band size: 80 kDa

false

• WB

Unknown

Western blot - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

Lanes 1-4 : Merged signal (red and green). Green - ab92324 observed at 80 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab92324 Anti-TNFAIP3 antibody [EPR2663] was shown to specifically react with TNFAIP3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265983 (knockout cell lysate ab257112) was used. Wild-type and TNFAIP3 knockout samples were subjected to SDS-PAGE. ab92324 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TNFAIP3 knockout HeLa cell lysate at 20 µg

Lane 3:

Jurkat cell treated with 5ng/ml PMA for 48 hours and then treated with 2µg/ml PHA for 48 hours, whole cell lysate at 20 µg

Lane 4:

Untreated Jurkat cell lysate at 20 µg

Predicted band size: 89 kDa

Observed band size: 80 kDa

false

• WB

Lab

Western blot - Anti-TNFAIP3 antibody [EPR2663] (AB92324)

All lanes:

Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/5000 dilution

Lane 1:

WEHI-3 (Mouse leukemia lymphoblast) whole cell lysate at 20 µg

Lane 2:

WEHI-3 treated with 20 ng/ml TNF alpha (<a href='/products/proteins-peptides/recombinant-human-tnf-alpha-protein-ab9642'>ab9642</a>) for 6 h 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

Predicted band size: 89 kDa

Observed band size: 80 kDa

false