All lanes : Anti-PGK1 antibody (ab90787) at 1 µg/mlLane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 2 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lysates/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 ab90787 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 ab90787 at 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 formaldehyde fixed (4%) (10 min) HeLa cells.
Anti-PGK1 antibody (ab90787) at 1 µg/ml + Human PGK1 full length protein (ab87630) at 0.1 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Exposure time : 30 seconds