All lanes : Anti-WSTF antibody [EPR1703] (ab109439) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : BAZ1B knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 171 kDa
Observed band size: 171 kDa

Lanes 1- 2: Merged signal (red and green). Green - ab109439 observed at 171 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

 ab109439 was shown to react with WSTF in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264907 (knockout cell lysate ab257370) was used. Wild-type HeLa and BAZ1B 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. ab109439 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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 : Anti-WSTF antibody [EP1704Y] (ab51256) at 1/15000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : BAZ1B knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 171 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab51256 observed at 175 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

 ab51256 was shown to react with WSTF in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264907 (knockout cell lysate ab257370) was used. Wild-type HeLa and BAZ1B 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. ab51256 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 15000 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.

Homozygous: 5 bp deletion in exon 2.