All lanes : Anti-Lamin A + Lamin C antibody [WL4G10] (ab232730) at 1/1000 dilution

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

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/20000 dilution

Predicted band size: 74 kDa
Observed band size: 70,75 kDa why is the actual band size different from the predicted?

Lanes 1-2: Merged signal (red and green). Green - ab232730 observed at 70,75 kDa. Red - loading control ab181602 observed at 37 kDa.

 ab232730 Anti-Lamin A + C antibody [WL4G10] was shown to specifically react with Lamin A + C in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261787 (knockout cell lysate ab256979) was used. Wild-type and Lamin A + C knockout samples were subjected to SDS-PAGE. ab232730 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 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.

 

All lanes : Anti-Lamin A + Lamin C antibody [4C11] (ab238303) at 1 µg/ml

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

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/20000 dilution

Predicted band size: 74 kDa
Observed band size: 74 kDa

Lanes 1-2: Merged signal (red and green). Green - ab238303 observed at 74 kDa. Red - loading control ab181602 observed at 37 kDa.

 ab238303 Anti-Lamin A + C antibody [4C11] was shown to specifically react with Lamin A + C in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261787 (knockout cell lysate ab256979) was used. Wild-type and Lamin A + C knockout samples were subjected to SDS-PAGE. ab238303 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 µg/ml 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: 19 bp deletion in exon 4.

 

Allele-2: 1 bp deletion in exon 4.

 

Allele-3: 22 bp deletion in exon 4.