All lanes : Anti-Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 antibody [20A] (ab2864) at 1/500 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CACNA2D1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 125 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?

Lanes 1-2: Merged signal (red and green). Green - ab2864 observed at 150 kDa. Red - loading control ab181602 observed at 37 kDa.  

 ab2864 Anti-Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 antibody [20A] was shown to specifically react with Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265080 (knockout cell lysate ab257374) was used. Wild-type and Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 knockout samples were subjected to SDS-PAGE. ab2864 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°CC at 1 in 500 Dilution and 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.

 

Allele-1: 14 bp deletion in exon 11.

 

Allele-2: 11 bp deletion in exon 11.

 

Allele-3: 1 bp insertion in exon 11.