A1806

Monoclonal Anti-Phosphotyrosine−Agarose antibody produced in mouse

clone PT-66, purified immunoglobulin, PBS solution

• Synonym(s): Monoclonal Anti-Phosphotyrosine, Phospho-Tyr, Phospho-tyrosine, p-Tyr
• UNSPSC Code: 12352203
• NACRES: NA.44

Properties

• biological source: mouse
• recombinant: expressed in mouse cell line
• conjugate: agarose conjugate
• antibody form: purified immunoglobulin
• antibody product type: primary antibodies
• clone: PT-66, monoclonal
• form: PBS solution
• technique(s): immunoprecipitation (IP): suitable
• isotype: IgG1
• capacity: 1 mg/mL binding capacity
• shipped in: wet ice
• storage temp.: 2-8°C
• target post-translational modification: phosphorylation (pTyr)

Description

General description

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 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from an immunized mouse.

Immunogen

phosphotyrosine conjugated to BSA

Application

Monoclonal Anti-Phosphotyrosine-Agarose antibody produced in mouse has been used in:

• immunoprecipitation experiments for affinity-purification of phosphotyrosine proteins
• phosphotyrosine pulldown assays
• PI3-kinase assay

Proteins containing phosphotyrosines were deteced in protein extracts from dissected tissue of Harlan Sprague Dawley rats or from transfected HEK293 cells using mouse monoclonal anti-phosphotyrosine antibody as the primary antibody. Reactivity was blocked when extracts were previously treated with 10 mM phosphotyrosine but not 10 mM tyrosine showing the specifity of the antibody for phosphorylated tyrosines. Immunoprecipitation of proteins containing phosphorylated tyrosines in rat dorsat root ganglion homogenates was performed using mouse monoclonal anti-phosphotyrosine. The antibody was incubated with the homogenates in the presence of 50 mm NaF and protease inhibitors for 4-14 hours at 4°. The antibody was precipitated using Protein G-agarose beads incubated overnight.

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 mediate 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 tyrosine kinase domains that undergo autophosphorylation when receptors bind to the respective ligands. Tyrosine-specific protein kinase activity has also been described in many retroviral oncogene proteins. Cells transformed by these oncogenes contain elevated levels of phosphotyrosine. Many of the oncogenes found in mammalian oncogenic viruses encode tyrosine protein kinases that reside in the cellular cytoplasm. Others encode transmembrane receptors whose tyrosine phosphotransferase activity is stimulated by the binding of ligand to the extracellular domain.

Physical form

Suspension in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide

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.

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 3
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable