All lanes : Anti-Thrombin Receptor antibody (ab63445) at 1/500 dilutionLane 1 : HeLa cell extracts treated with Nocodazole (1ug/ml, 18hours)Lane 2 : HeLa cell extracts treated with Nocodazole (1ug/ml, 18hours) with immunising peptide at 10 µgLysates/proteins at 30 µg per lane.
ICC/IF image of ab63445 stained Hek293 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 (ab63445, 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.