ICC/IF image of ab41437 stained HepG2 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 ab41437 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
Lanes 1 - 2 : anti-SHP1 (C-terminal) Lanes 3 - 5 : Anti-SHP1 (phospho Y536) antibody (ab41437) at 1/1000 dilutionLane 1 : human Jurkat cells treated with 1 mM pervanadate for 30 min. Lane 2 : human Jurkat cells treated with 1 mM pervanadate for 30 min, blot exposed to alkaline phosphatase before probing Lane 3 : human Jurkat cells treated with 1 mM pervanadate for 30 min. Lane 4 : human Jurkat cells treated with 1 mM pervanadate for 30 min, blot exposed to alkaline phosphatase before probing Lane 5 : human Jurkat cells treated with 1 mM pervanadate for 30 min. with phospho-Tyr536-SHP1 peptide