Phospho Threonine monoclonal antibody, clone 18F6 (Cat # MAB2001) is shown to detect threonine phosphorylation of proteins in a lysate from A-431 cells stimulated with EGF. Separation is achieved under reducing conditions using a pre-cast 4-20% iGel from Gradipore, Inc. A 1 : 1,000 dilution of Phospho Threonine monoclonal antibody, clone 18F6 (Cat # MAB2001) is used for 30 min followed by detection using a 1 : 2,500 dilution of IRDye® 800 conjugated Goat-a-Mouse IgG [H&L] and visualization using the Odyssey® Infrared Imaging System developed by LI-COR. Other detection systems will yield similar results. IRDye is a trademark of LI-COR, Inc.
ELISA results of PhosphoThreonine monoclonal antibody, clone 18F6 (Cat # MAB2001) tested against BSA conjugates of pT, pY and pS. Each well was coated with 0.1 ug of conjugate. The starting dilution of antibody was 1 : 1000 and each point on the X-axis represents a 2-fold dilution.