All lanes : Anti-HSF1 antibody (ab105086) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 3 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 6 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate Lane 7 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab105086 stained MCF7 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 (ab105086, 1µg/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. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa, Hek293 and HepG2 cells at 1µg/ml.