All lanes : Anti-N myc interactor/NMI antibody [EPR11065(2)] (ab183724) at 1/1000 dilution

Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : NMI knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 35 kDa
Observed band size: 39 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab183724 observed at 39 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab183724 Anti-N myc interactor/NMI antibody [EPR11065(2)]  was shown to specifically react with N myc interactor/NMI in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267013 (knockout cell lysate ab258077) was used. Wild-type and N myc interactor/NMI knockout samples were subjected to SDS-PAGE. ab183724 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: 2 bp deletion in exon5

 

Allele-2: 330 bp insertion in exon 5.