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CEA / CEACAM-5 Antibody (MIC0101) in WB
Western blot analysis of Carcinoembryonic Antigen (CEA) was performed by loading 20ug of BxPC-3, HeLa, and HepG2 whole cell lysates per well and 5ul of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. CEA (with varying degrees of glycosylation) was detected at ~77-180kD using a CEA monoclonal antibody (Product # MIC0101) at a dilution of 1ug/ml in 5% milk in TBST overnight at 4C on a rocking platform, followed by a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico substrate (Product # 34080).
