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Human IgA (Alpha heavy chain) Antibody (MA5-11208) in WB
Western blot analysis of IgA was performed by loading 0.5ug of purified Human IgA, Human IgG, Human IgM, and Mouse IgA, and 10ul of PageRuler Prestained Protein Ladder (Product ## 26616) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membranes using the G2 Fast Blotter (Product ## 62288), and blocked with 5% BSA in TBST for at least 1 hour at room temperature. IgA heavy chain was detected at ~55kD using a mouse anti-human IgA monoclonal antibody (Product ## MA5-11208) diluted to a concentration of 0.5ug/ml in blocking buffer, overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse light chain secondary antibody diluted 1:40,000 for at least 30 minutes at room temperature (panel A). To verify the presence of all isotypes, a second blot was probed with a goat anti-human IgM + IgG +IgA polyclonal antibody (Product ## 31128) at a concentration of 0.5ug/ml in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated mouse anti-goat IgG secondary antibody (Product ## 31400) diluted 1:40,000 in blocking buffer for at least 30 minutes at room temperature (panel B). Chemiluminescent detection was performed using SuperSignal West Pico.
