The cellular phosphotyrosine direct enzyme-linked immunoabsorbant assay (ELISA) kit can be used to characterize tyrosine phosphorylation in adherent cultured cells. It has been shown to be useful for analyzing the effect of inhibitors of cellular tyrosine phosphorylation. This kit requires a model cell line provided by the researcher that can be stimulated in vitro to undergo tyrosine phosphorylation. Human A431 carcinoma cell and foreskin fibroblast cells stimulated with epidermal growth factor as well as mouse 3T3 fibroblasts stimulated with insulin have been successfully used with this kit.Applications:Characterization of tyrosine phosphorylation in adherent cultured cells Capacity:192 Assays Components: C2609-50A: Phosphotyrosine (HRP) 1x200ugC2609-50B: Blocking Reagent 1x2gC2609-50C: TMB tablets 1x4tabletsC2609-50D: Phosphate-Citrate Buffer (5x) 1x8mlC2609-50E: DMSO 1x4mlC2609-50F: Tissue culture plates 2x96wellsOther components required but not included as part of kit are:Fresh Hydrogen Peroxide 30% Model cell lineCulture mediaGrowth Factor or cytokinePhosphate Buffered Saline (PBS), pH 7.4Protein tyrosine kinase inhibitorsELISA Plate Reader with filter for single wavelength for 375nm, 450nm, and/or 655nm37°C incubatortimervariable volume (5-200l) pipet + tips5, 10 and 25ml pipetsAcetic acidMethanol0.5M H2SO4Storage and Stability:Stable for 6 months at 4°C from date of shipment.
