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A5964 Sigma

Monoclonal Anti-Phosphotyrosine–Peroxidase antibody produced in mouse

clone PT-66, purified immunoglobulin, lyophilized powder

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, Antibodies to Phosphoproteins,
biological source   mouse
antibody form   purified immunoglobulin
clone   PT-66, monoclonal
form   lyophilized powder
packaging   vial of 0.2 mL conjugate
application(s)   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
isotype   IgG1
conjugate   peroxidase conjugate
storage temp.   2-8°C

Description

Immunogen

phosphotyrosine conjugated to BSA

Physical form

Lyophilized from a solution containing 1% bovine serum albumin and 0.01% thimerosal in 0.01 M sodium phosphate buffered saline.

Application

Affnitiy purification of proteins from ram and bear spermatozoa was performed using agarose-linked anti-phosphotyrosine antibody with incubation for 2 hours at 4 °C.

Monoclonal anti-phosphotyrosine–peroxidase antibody can be used in dot blot (diluted 1:40,000) and chemiluminescence dot blot (diluted 1:200,000) using phosphotyrosine-BSA. It can also be used in direct ELISA (diluted 1:60,000).

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 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. 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.

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RIDADR 
NONH for all modes of transport
WGK Germany 
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Keywords: Amplification, Buffers, Cell biology, Enzyme-linked immunosorbent assay, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Molecular biology, Phosphorylations, Purification, Western blot

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