All lanes : Anti-PKM2 antibody (ab85555) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : MEL-1 (Human embryonic stem cell, male cell line) Whole Cell Lysate (ab27198)Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 4 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 5 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate Lane 6 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate Lane 7 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab85555 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 (ab85555, 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. This antibody also gave a positive result in 100% Methanol fixed (5 min) HepG2 cells at 5µg/ml, and in 4% PFA fixed (10 min) HeLa, and HepG2 cells at 5µg/ml.
ab85555 staining PKM2 in HeLa cells treated with resveratrol (ab120726), by ICC/IF. Decrease in PKM2 expression correlates with increased concentration of resveratrol as described in literature.The cells were incubated at 37°C for 48h in media containing different concentrations of ab120726 (resveratrol) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab85555 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.