Anti-TNFAIP3 antibody [59A426]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-TNFAIP3 antibody [59A426] (AB13597)

Overlay histogram showing HepG2 cells stained with ab13597 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13597, 2μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HepG2 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNFAIP3 antibody [59A426] (AB13597)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human placenta tissue labelling TNFAIP3 with ab13597 at 5μg/ml. Staining was enhanced by boiling tissue sections in 10mM sodium citrate buffer, pH6.0 for 10-20 minutes followed by cooling at room temperature for 20 minutes.

• WB

Lab

Western blot - Anti-TNFAIP3 antibody [59A426] (AB13597)

Lanes 1-4 : Merged signal (red and green). Green - ab13597 observed at 80 kDa. Red - loading control, ab181602 observed at 37 kDa.

ab13597 Anti-TNFAIP3 antibody [59A426] 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. ab13597 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597) at 1/500 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='/ko/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

Supplier Data

Western blot - Anti-TNFAIP3 antibody [59A426] (AB13597)

Lanes 1-4 : Merged signal (red and green). Green - ab13597 observed at 80 kDa. Red - loading control, ab181602 observed at 37 kDa.

ab13597 Anti-TNFAIP3 antibody [59A426] 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. ab13597 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597) at 1/500 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

Supplier Data

Western blot - Anti-TNFAIP3 antibody [59A426] (AB13597)

All lanes:

Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597) at 4 µg/mL

All lanes:

Jurkat cell lysate

Predicted band size: 89 kDa

false

• WB

Unknown

Western blot - Anti-TNFAIP3 antibody [59A426] (AB13597)

All lanes:

Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597) at 1 µg/mL

Lane 1:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 2:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 89 kDa

Observed band size: 15 kDa,34 kDa,90 kDa

true

Exposure time: 20min