This antibody recognizes Thymidylate Synthase and an additional band corresponding to the ternary complex of hTS-dFUMP-reduced folate in HeLa cells treated with the TS inhibitor 5-FUdR. (This event occurs in most human breast, colorectal, gastric, head and neck carcinomas.) The antibody can detect Thymidylate Synthase by immunochemistry in proliferating cell cultures and tissues but does not stain nonproliferating cells. Normal colon mucosa shows weak staining; however, some colorectal cancer specimens show very strong staining.
ICC/IF image of ab34836 stained HeLa 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 (ab34836, 5µg/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.