All lanes : Anti-STAT1 antibody [EPR4407] (ab109320) at 1/10000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : STAT1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 87 kDa
Observed band size: 87 kDa

Lanes 1- 2: Merged signal (red and green). Green - ab109320 observed at 87 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

 ab109320 was shown to react with STAT1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255346 (knockout cell lysate ab263837) was used. Wild-type HeLa and STAT1 knockout HeLa 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. ab109320 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 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.

Allele-1: 14 bp deletion in exon 4.

Allele-2: 5 bp deletion in exon 4.

Allele-3: 1 bp deletion in exon 4.