All lanes : Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : STK4 knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : Ramos cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution

Predicted band size: 55 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab51134 observed at 52 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab51134 Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] was shown to specifically react with MST-1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265442 (knockout cell lysate ab258215) was used. Wild-type and MST-1 knockout samples were subjected to SDS-PAGE. ab51134 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.

 

Allele-1: 67 bp insertion in exon 3.

 

Allele-2: Insertion of the selection cassette in exon 3.