All lanes : Anti-Rab5b antibody (ab72907) at 1 µg/mlLane 1 : Placenta (Human) Tissue Lysate - adult normal tissue (ab29745)Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 6 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab72907 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 (ab72907, 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.