ICC/IF image of ab73998 stained HCT116 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 (ab73998, 5µg/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-Phospholipase C beta 3 (phospho S537) antibody (ab73998) at 1/500 dilutionLane 1 : Extracts from Jurkat cells, treated with UV (15mins)Lane 2 : Extracts from Jurkat cells, treated with UV (15mins) with immunising phosphopeptide at 10 µgLysates/proteins at 30 µg per lane.
Immunohistochemistry analysis of paraffin-embedded human brain tissue using 1/50 ab73998. Left: untreated sample. Right: sample treated with immunising phosphopeptide.