All lanes : Anti-CF150 Antibody (Center) at 1:1000 dilution Lane 1: human cerebellum lysates Lane 2: human spleen lysates Lane 3: HT-29 whole cell lysates Lane 4: MCF-7 whole cell lysates Lane 5: THP-1 whole cell lysates Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/10000 dilution Predicted band size : 59 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
Overlay histogram showing U-2OS cells stained with AW5397 (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 (AW5397, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(NA168821) at 1/400 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.