All lanes : Anti-Sp5 antibody (ab36593) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell LysateLane 4 : HEK293 Human embryonic kidney cell line Whole Cell LysateLane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell LysateLane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell LysateLane 8 : U2OS (Human osteosarcoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.
ICC/IF image of ab36593 stained human HEK 293 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab36593, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa and MCF7 cells.