ICC/IF image of ab51136 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab51136, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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
All lanes : Anti-Hsp90 beta (phospho S254) antibody (ab51136) at 1/500 dilutionLane 1 : extracts from Hela cells treated with TNF-a with immunizing peptideLane 2 : extracts from Hela cells treated with TNF-aLysates/proteins at 0.02 µg/ml per lane.
Ab51136 at 1/50 dilution staining human breast carcinoma tissue with and without blocking peptide; paraffin embedded.Ab51136 at 1/50 dilution staining human breast carcinoma tissue with and without blocking peptide; paraffin embedded.
All lanes : Anti-Hsp90 beta (phospho S254) antibody (ab51136) at 1/1000 dilutionLane 1 : Mouse heart whole tissue lysateLane 2 : Mouse heart whole tissue lysateLysates/proteins at 50 µg per lane.SecondaryAn HRP-conjugated Goat anti-rabbit polyclonal. at 1/5000 dilution