All lanes : Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution

Lane 1 : Wild-type A549 cell lysate
Lane 2 : DDX58 knockout A549 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 107 kDa
Observed band size: 107 kDa

Lanes 1 - 2: Merged signal (red and green). Green - ab180675 observed at 107 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab180675 was shown to react with DDX58 in A549 wild-type cells in western blot with loss of signal observed in DDX58 knockout cell line ab267117 (DDX58 knockout cell lysate ab257917). Wild-type and DDX58 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab180675 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Allele-1: 16 bp deletion in exon2

 

Allele-2: 8 bp deletion in exon 2.

 

Allele-3: 5 bp deletion in exon 2.