Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor® Plus 800 conjugate for Western Blot
Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody (A32730) in WB
Western blot analysis of protein disulphide-isomerase (PDI) and progesterone receptor complex (p23) was performed by loading 3-fold serial dilutions of A431 whole cell lysate (starting at 15ug) and 2ul of the PageRuler Prestained NIR Protein Ladder (Product # 26635) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to Nitrocellulose Membranes (Product # 88018) and blocked with Fluorescence Blocker for 30 min. Membranes were probed with a PDI monoclonal antibody (Product # MA3-019) at a dilution of 1:5000 and a p23 monoclonal antibody at a dilution of 1:1000 (Product # MA3-414) overnight at 4°C on a rocking platform, washed with TBST, and probed with an Invitrogen Alexa Fluor Plus 800 Goat anti-Mouse IgG secondary antibody (Product # A32730) at dilutions of 1:40,000 for 45 minutes. Blots were imaged on an Infrared fluorescence imaging system.