All lanes : Anti-PKC delta antibody [EPR17075] (ab182126) at 1/5000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : PRKCD knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 78 kDa
Observed band size: 78 kDa

Lanes 1 - 2: Merged signal (red and green). Green - ab182126 observed at 78 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab182126 was shown to react with PKC in wild-type HEK-293T cells in western blot. The bands observed in PRKCD knockout cell line ab266143 (PRKCD knockout cell lysate ab257042) below 78kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and PRKCD knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab182126 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4

176;

>C at a 1 in 5000 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: 2 bp deletion in exon2

 

Allele-2: 1 bp insertion in exon 2.