direct ELISA: 1:60,000 using Phosphotyrosine-BSA dot blot: 1:40,000-1:200,000 using phosphotyrosine-BSA using chromogenic and chemiluminescent substrates, respectively
As determined by ELISA and competitive ELISA, the antibody reacts specifically with phosphorylated tyrosine, both as free amino acid or conjugated to carriers such as BSA or KLH. No cross-reactivity is observed with non-phosphorylated tyrosine, phosphothreonine, phosphoserine, AMP or ATP.
Monoclonal Anti-Phosphotyrosine (mouse IgG1 isotype) is derived from the PT-66 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from BALB/c mice immunized with a phosphotyrosine-BSA conjugate.
Specificity
Mouse anti-phosphotyrosine-peroxidase antibody reacts specifically with phosphorylated tyrosine coupled to BSA. The product has shown no reactivity for other phosphorylated amino acids like phosphothreonine and phosphoserine.
Immunogen
phosphotyrosine conjugated to BSA
Application
Monoclonal Anti-Phosphotyrosine-Peroxidase antibody has been used in
enzyme linked immunosorbent assay (ELISA)
dot blot
Chemiluminescence dot blot
kinase assay
Biochem/physiol Actions
Reversible phosphorylation of proteins is an important post-translational modification that plays a regulatory role in the expression of most proteins in the cells. Reversible phosphorylation at multiple serine, tyrosine and threonine residues mediates numerous signalling pathways in both prokaryotic and eukaryotic cells . Cellular proteins with phosphorylated tyrosine increase many fold by the activation of tyrosine kinases. Most mitogenic receptor systems such as EGF, PDGF, insulin receptors contain serine/threonine/tyrosine kinase domains that undergo autophosphorylation when receptors bind to the respective ligands. Monoclonal anti-phosphotyrosine–peroxidase antibody can be used in kinase assay to visualize the fraction of phosphorylated substrate.
Physical form
Lyophilized from a solution containing 1% bovine serum albumin and 0.05% MIT in 0.01 M sodium phosphate buffered saline.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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