All lanes : Anti-ICAM1 antibody [EP1442Y] (ab53013) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ICAM1 knockout HeLa cell lysate
Lane 3 : Raji 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 1/10000 dilution

Predicted band size: 57 kDa

Lanes 1-4: Merged signal (red and green). Green - ab53013 observed at 90 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab53013 Anti-ICAM1 antibody [EP1442Y]  was shown to specifically react with ICAM1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261742 (knockout cell lysate ab256947) was used. Wild-type and ICAM1 knockout samples were subjected to SDS-PAGE. ab53013 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: Insertion of the selection cassette in exon 2.

 

Allele-2: 1 bp insertion in exon 2.