All lanes : Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/1000 dilution

Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : TNFAIP3 knockout A549 (Human lung carcinoma cell line) whole cell lysate
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
Lane 4 : Untreated Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 89 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?

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.

Homozygous: 10 bp deletion in exon7