All lanes : Anti-WHSC1/NSD2 antibody [29D1] (ab75359)

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : WHSC1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 152 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab75359 observed at 80 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

 ab75359 was shown to react with WHSC1/NSD2 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264936 (knockout cell lysate ab257798) lane below 80kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and WHSC1 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. ab75359 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at a 5 µg/ml and a 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 20000 dilution for 1 hour at room temperature before imaging.

Homozygous: 1 bp insertion in exon 2.