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Estrogen Receptor Antibody (MA5-14598) in WB
Western blot analysis of Estrogen Receptor (ER) was performed by loading the indicated amounts of recombinant full-length human ER alpha (Product ## A15674) and ER beta (Product ## A15664) proteins, and 10ul of PageRuler Prestained Protein Ladder (Product ## 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product ## 88518) using the G2 Blotter (Product ## 62288), and blocked with 5% BSA in TBST for at least 1 hour at room temperature. ER alpha was detected at ~66kD (left panel) using an ER monoclonal antibody (Product ## MA5-14598) at a dilution of 1:200 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG secondary antibody (Product ## 31430) at a dilution of 1:40,000. Chemiluminescent detection was performed using SuperSignal West Pico (Product ## 34080). Directly alongside the above blot, a second blot containing the same recombinant ER alpha and ER beta protein samples was probed with an ER-beta specific polyclonal antibody (Product ## PA1-313, right panel) at a dilution of 1:2000 overnight at 4C, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product ## 31460) at a dilution of 1:40,000. Chemiluminescent detection was also performed using SuperSignal West Pico (Product ## 34080).
