Overlay histogram showing Jurkat cells stained with BD-PB4116 (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody for 60 min at 37ºC. The secondary antibody used was Goat-Anti-mouse IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed) at 1/400 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was mouse IgG (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

All lanes : Anti-MGMT Antibody at1:1000-1:2000 dilution Lane 1: Jurkat whole cell lysate Lane 2: MCF-7 whole cell lysate Lane 3: MOLT-4 whole cell lysate Lane 4: Raji whole cell lysate Lysates/proteins at 20 μg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 22 kDa Blocking/Dilution buffer: 5% NFDM/TBST.