All lanes : Anti-p53R2 antibody [EPR8816] (ab154194) at 1/1000 dilution

Lane 1 : Wild-Type HCT116 cell lysate
Lane 2 : RRM2B knockout HCT116 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 40 kDa
Observed band size: 40 kDa

Lanes 1- 2: Merged signal (red and green). Green - ab154194 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

 ab154194 was shown to react with p53R2 in Wild-Type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266897 (knockout cell lysate ab257216) was used. Wild-Type HCT116 and RRM2B knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154194 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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: 1 bp insertion in exon1

 

Allele-2: Insertion of the selection cassette in exon 1.