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  • A1806 - Monoclonal Anti-Phosphotyrosine−Agarose antibody produced in mouse

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A1806 Sigma

Monoclonal Anti-Phosphotyrosine−Agarose antibody produced in mouse

clone PT-66, purified immunoglobulin, PBS solution

Synonym: Monoclonal Anti-Phosphotyrosine, Phospho-Tyr, Phospho-tyrosine, p-Tyr

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Properties

Related Categories Alphabetical Index, Antibodies, Antibodies for Cell Biology, Antibodies for Kinase/Phosphatase Biology, Capture Media for Phosphoproteins,
biological source   mouse
recombinant   expressed in mouse cell line
antibody form   purified immunoglobulin
clone   PT-66, monoclonal
form   PBS solution
application(s)   immunoprecipitation: suitable
isotype   IgG1
capacity   1 mg/mL binding capacity
conjugate   agarose conjugate
shipped in   wet ice
storage temp.   2-8°C

Description

Immunogen

phosphotyrosine conjugated to BSA

Physical form

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

Application

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.

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.

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.

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Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

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