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Merck

A1806

Sigma-Aldrich

Monoclonal Anti-Phosphotyrosine−Agarose antibody produced in mouse

clone PT-66, purified immunoglobulin, PBS solution

Sinónimos:

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

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.44

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)

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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.

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.

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Storage Class

10 - Combustible liquids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable


Certificados de análisis (COA)

Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»

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B P Ceresa et al.
Molecular and cellular biology, 18(7), 3862-3870 (1998-06-25)
To examine the role of clathrin-dependent insulin receptor internalization in insulin-stimulated signal transduction events, we expressed a dominant-interfering mutant of dynamin (K44A/dynamin) by using a recombinant adenovirus in the H4IIE hepatoma and 3T3L1 adipocyte cell lines. Expression of K44A/dynamin inhibited
RET oncoproteins induce tyrosine phosphorylation changes of proteins involved in RNA metabolism
Gorla L, et al.
Cellular Signalling, 18(12), 2272-2282 (2006)
Insulin activation of phosphatidylinositol 3-kinase in human skeletal muscle in vivo
Hickey MS, et al.
Journal of Applied Physiology, 83(3), 718-722 (1997)
Tyrosine 116 of the herpes simplex virus type 1 IE alpha 22 protein is an ocular virulence determinant and potential phosphorylation site
Brandt CR and Kolb AW
Investigative Ophthalmology & Visual Science, 44(11), 4601-4607 (2003)
Helena L Palka et al.
The Journal of biological chemistry, 278(8), 5728-5735 (2002-12-12)
The receptor protein-tyrosine phosphatase (PTP) DEP-1 (CD148/PTP-eta) has been implicated in the regulation of cell growth, differentiation, and transformation, and most recently has been identified as a potential tumor suppressor gene mutated in colon, lung, and breast cancers. We have

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