All lanes : Anti-Endo G antibody (ab64668) at 1 µg/mlLane 1 : Skeletal Muscle (Mouse) Tissue Lysate (ab29711)Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 4 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate - tumor cell line (ab30124)Lane 5 : Liver (Mouse) Tissue Lysate - normal tissueLane 6 : Rat Kidney Tissue LysateLane 7 : THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate (ab7913)Lysates/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 ab64668 stained HeLa cells. The cells were fixed with 100% Methanol (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 (ab64668 at 1µg/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). This antibody also gave a positive result in 100% Methanol (5 min), and 4% Pfa (10minutes) in MCF-7, Hek293 and HepG2 cells at 1µg/ml.
ab64668 (1/2000) staining Endo G in paraffin-embedded Human pancreas tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. The secondary antibody was goat anti rabbit/mouse conjugated to HRP. For further experimental details please refer to abreview.See Abreview