All lanes : Anti-Hsp90 beta antibody (ab51145) at 1/500 dilutionLane 1 : extracts from Hela cells treated with TNF-aLane 2 : extracts from Hela cells treated with TNF-a with immunizing peptideLysates/proteins at 0.02 µg/ml per lane.
Ab51145 at 1/50 dilution staining Human breast carcinoma, with and without blocking peptide; paraffin embedded.
ICC/IF image of ab51145 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51145, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.