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P3300

Sigma-Aldrich

Monoclonal Anti-Phosphotyrosine antibody produced in mouse

clone PT-66, ascites fluid

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

biological source

mouse

Quality Level

conjugate

unconjugated

antibody form

ascites fluid

antibody product type

primary antibodies

clone

PT-66, monoclonal

contains

15 mM sodium azide

technique(s)

indirect ELISA: 1:1,600
western blot: 1:2,000 using a fresh human platelet preparation

isotype

IgG1

shipped in

dry ice

storage temp.

−20°C

target post-translational modification

unmodified

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This Item
P5872P3555P1869
biological source

mouse

biological source

mouse

biological source

mouse

biological source

-

antibody form

ascites fluid

antibody form

purified immunoglobulin

antibody form

ascites fluid

antibody form

purified immunoglobulin

conjugate

unconjugated

conjugate

unconjugated

conjugate

unconjugated

conjugate

unconjugated

isotype

IgG1

isotype

IgG1

isotype

IgG2b

isotype

IgG2b

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

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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.
Phosphorylation, attachment of a phosphate group to protein, alters protein functionality by activation or deactivation of the protein, in which protein kinase plays a crucial role. Tyrosine residue phosphorylation plays a key role in cell growth and differentiation. The monoclonal anti-phosphotyrosine antibody is useful in immunoblotting for identification of phosphotyrosine containing protein from cultured human epidermoid carcinoma cell line A-431 and human platelets preparation. This product can be used in immunofluorescent labeling of tyrosine residue at focal adhesion and cellular junctions of cultured MDCK cells. This product is also useful in studies of signal transduction and growth factor receptors. This antibody has shown specificity for binding to proteins that contain phosphorylated tyrosine residue either in free amino acid form or in conjugation with KLH or BSA but will not react with non-phosphorylated tyrosine or other phosphorylated proteins and amino acids. It will also not react with phosphorylated molecules like ATP or AMP. Monoclonal anti-phosphotyrosine reacts specifically with mouse and human.

Specificity

This antibody is specific for phosphorylated tyrosine both as the free amino acid or when conjugated to carriers such as BSA or KLH

Immunogen

phosphotyrosine conjugated to BSA

Application

Monoclonal Anti-Phosphotyrosine antibody produced in mouse has been used in
  • immunocytochemistry
  • immunoprecipitation
  • western blotting
  • peptide binding studies flow cytometry
  • enzyme linked immunosorbent assay (ELISA)
  • radioimmunoassay (RIA)
  • immunoaffinity isolation

Monoclonal anti-phosphotyrosine antibody produced in mouse is used in immunohistochemisty, flow cytometry, immunoprecipitation, immunoblotting, ELISA and RIA for localization of phosphorylated tyrosine containing proteins. It can also be used for immunoaffinity isolation.

Biochem/physiol Actions

Phosphotyrosine levels enhanced in cellular proteins leads to activation of several cellular processes mediated by phosphotyrosine kinases. Phosphotyrosine residues that are autophosphorylated by the binding of ligands associated with receptors like epidermal growth factor (EGF), platelet-derived growth factor (PDGF) and insulin receptors, functions as the primary integral component of mitogenic signaling cascade. Phosphotyrosine mediates the activation of T-cell as a result of phosphorylation of tyrosine residues by tyrosine kinases in cytoplasmic domains of CD4 an CD8 which in turn phosphorylates TCR-CD3 complex. Phosphotyrosine activity has also been found in many retroviruses whose oncogenes encodes tyrosine-specific protein kinase.
Tyrosine residue phosphorylation plays a key role in cell growth and differentiation.

Physical form

This product is supplied as ascites fluid containing 15 mM sodium azide

Storage and Stability

For continuous use, store at 2-8 °C for up to one month. For extended storage, the solution may be frozen in working aliquots. Repeated freezing and thawing is not recommended. Storage in "frost-free" freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use.

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.

related product

Product No.
Description
Pricing

Storage Class

13 - Non Combustible Solids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


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T R Cimato et al.
The Journal of cell biology, 138(5), 1089-1103 (1997-09-08)
Protein methylation is a posttranslational modification that can potentially regulate signal transduction pathways in a similar manner as protein phosphorylation. The role of protein methylation in NGF signaling was examined by metabolic labeling of PC12 cell proteins with L-[methyl-3H]methionine and
Insertional oncogenesis by non-acute retroviruses: implications for gene therapy
Fan H and Johnson C
Viruses, 3(4), 398-422 (2011)
S Barlati et al.
FEBS letters, 281(1-2), 137-140 (1991-04-09)
Immunoblotting analysis of purified human urokinase plasminogen activator (u-PA), gives a positive signal when reacted with anti-phosphotyrosine monoclonal antibodies (MoAb anti-P-Tyr); competition with o-phospho-DL-tyrosine (P-Tyr) but not o-phospho-DL-threonine or serine (P-Treo, P-Ser) completely suppresses this signal. Either the 55 kDa
Nicholas G Clarkson et al.
The Journal of biological chemistry, 284(37), 24725-24734 (2009-07-10)
Regulation by the NK and T cell surface receptor CD244 in mice and humans depends both on engagement at the cell surface by CD48 and intracellular interactions with SAP and EAT-2. Relevance to human disease by manipulating CD244 in mouse
Epidermal Growth Factor Receptor Down-Regulation Triggers Human Myoblast Differentiation
Boisguerin P, et al.
Chembiochem, 8(18), 2302-2307 (2007)

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