Anti-Eck (EphA2, Mpk-5, Native Epithelial Cell Kinase), clone 0.T.34

Applications: , Suitable for use in Western Blot, Protein Kinase Assay, Immunoprecipitation and Immunocytochemistry. Other applications not tested. Recommended Dilution: Western Blot: 1-2ug/ml, Detected Eck in RIPA lysates from human A431 cells and previously from foreskin fibroblasts, murine 3T3/A31 and rat L6 cells. A431 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with E3360-01B. Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system, Protein Kinase Assay: Reported to have been used in an IP autophosphorylation assay, using a Mn-PIPES reaction buffer., Immunoprecipitation: Reported to immunoprecipitate Eck from 500ug of a human breast epithelial cell line which had been lysed in TBS containing 1% Triton X-100. Use 1-4ug per reaction., Immunocytochemistry: Reported to immunostain Eck in human, mouse and rat epithelial cells fixed with 3.7% formaldehyde solution and permeabilized with 0.5% Triton X-100 in TBS., Optimal dilutions to be determined by the researcher. Recommended Secondary Antibody: I1904-77B: IgG1 (HRP) Pab Gt x Mo, Positive Antigen Control: A0001-50: Non-stimulated A431 lysate, Control cells: CHO cell lysate, foreskin fibroblasts, 3T3/A31 cells, L6 cells. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.