Phospho Tyrosine monoclonal antibody, clone 13F9 (Cat # MAB2002) is shown to detect by immunoblot (Lane 1) phosphorylated EGFR in an A-431 cell lysate (10 ug per lane) after stimulation with EGF. The binding of EGF to EGFR causes rapid activation of intrinsic autophosphorylation of multiple tyrosine residues in the cytoplasmic domain of the protein. This propagates the ERK1/2 signal pathway which regulates cell growth, survival, proliferation and differentiation. Lane 2 shows total protein in the lysate after coommassie staining. Minimal reactivity occurs when the antibody is used to stain a lysate from unstimulated A-431 cells. A 4-20% gradient gel was used for separation prior to transfer to nitrocellulose. A 1 : 1,000 dilution of Phospho Tyrosine monoclonal antibody, clone 13F9 (Cat # MAB2002) is used at room temperature for 60 min followed by detection using IRDye™800 Conjugated Goat-a-Mouse IgG [H&L] diluted 1 : 2,500 for 30' at room temperature. Image w
ELISA results of PhosphoTyrosine monoclonal antibody, clone 13F9 (Cat # MAB2002) 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. HRP conjugated Goat-anti-Mouse IgG H&L and TMB substrate were used for detection.