Western Blot; Positive WB detected in: Hela whole cell lysate, MCF-7 whole cell lysate, K562 whole cell lysate, HL-60 whole cell lysate, HepG2 whole cell lysate, A549 whole cell lysate, Jurkat whole cell lysate, PC3 whole cell lysate, Rat brain tissue; All lanes: Hsp90 alpha + beta antibody at 1.25µg/ml; Secondary; Goat polyclonal to rabbit IgG at 1/50000 dilution; Predicted band size: 90 KDa; Observed band size: 90 KDa;
Immunoprecipitating Hsp90 alpha + beta in Hela whole cell lysate; Lane 1: Rabbit control IgG instead in Hela whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000); Lane 2: RAC07044 (3µg) + Hela whole cell lysate (500µg); Lane 3: Hela whole cell lysate (20µg);
Overlay histogram showing Jurkat cells stained with RAC07044 (red line) at 1:50. The cells were fixed with 70% Ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4?. The secondary antibody used was FITC goat anti-rabbit IgG (H+L) at 1/200 dilution for 1 h at 4?. Control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.