All lanes : Anti-STAT5b (phospho S731) antibody (ab52211) at 1/500 dilutionLane 1 : Extracts from RAW264.7 cells treated with EGF (200ng/ml, 30min) Lane 2 : Extracts from RAW264.7 cells treated with EGF (200ng/ml, 30min), ab52211 was pre-incubated with immunising (blocking) peptide
Paraffin-embedded human breast carcinoma tissue stained for STAT5b (phospho S731), using ab52211 (1/100). The right hand panel represents a negative control where ab52211 was pre-incubated with the immunising (blocking) peptide.
ICC/IF image of ab52211 stained HeLa cells. The cells were 4% PFA 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 (ab52211, 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.