All lanes : Anti-MADH7 antibody (ab76498) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab52255)Lane 3 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab46770)Lysates/proteins at 10 µg per lane.SecondaryAnti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) at 1/50000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab76498 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 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 ab76498 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 HeLa, Hek293, and MCF-7 cells in 4% formaldehyde (10min) at 5ug/ml, and in HeLa, Hek293, HepG2, and MCF-7 cell lines in 100% Methanol (5min) at 5ug/ml.
ICC/IF images of ab76498 stained mouse embryonic stem cells. The cells were fixed in 4% Paraformaldehyde, permeabilized using 0.1% Triton X-100 and blocked with 1% Goat serum, 0.1% BSA in PBS for 30 minutes at RT, before incubation with ab76498 at a 1/200 dilution for 2 hours at RT. The secondary used was an Alexa Fluor 488 conjugated goat anti-Rabbit polyclonal at a 1/200 dilution. 1) D3 mouse ES cells stained with anti-MADH7 (ab76498). 2) DAPI staining for image 1. 3) Ecad-/- mES cells (derived from D3 Mouse Embryonic Stem Cell line) stained with anti-MADH7 antibody (ab76498). 4) DAPI staining for image 3.See Abreview