All lanes : Anti-67kDa Laminin Receptor antibody (ab99484) at 1 µg/mlLane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lane 2 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate Lane 3 : HCT 116 (Human Colorectal Carcinoma) Whole Cell Lysate Lane 4 : LOVO (Human colon adenocarcinoma cell line) Whole Cell LysateLane 5 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate Lane 6 : HeLa (Human epithelial carcinoma 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 ab99484 stained HepG2 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 ab99484 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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 methanol fixed (100%, 5min) HeLa, HEk293, and MCF-7 cells at 1ug/ml, and in formaldehyde fixed (4%, 10min) HeLa, HEk293, HepG2, and MCF-7 cells at 1ug/ml.