Allele-1: 31 bp deletion in exon 2.

 

All lanes : Anti-Yes1 antibody [EPR3173] (ab109265) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : YES1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 60 kDa
Observed band size: 60 kDa

Lanes 1- 4: Merged signal (red and green). Green - ab109265 observed at 60 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

 ab109265 was shown to react with Yes1 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab265202 (knockout cell lysate ab258290) lane below 60kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and YES1 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. ab109265 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated 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.

Allele-2: 14 bp deletion in exon 2.

 

Allele-3: 8 bp deletion in exon 2.