单克隆抗-磷酸酪氨酸

NA.44

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.
经 ELISA 和竞争性 ELISA 确定，该抗体与磷酸化的酪氨酸特异性反应，该磷酸化的酪氨酸既可以是游离氨基酸形式，也可与 BSA 或 KLH 等载体结合的形式。与非磷酸化的酪氨酸、磷酸苏氨酸、磷酸丝氨酸、AMP 或 ATP 没有观察到交叉反应。

与 BSA 结合的磷酸酪氨酸

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.

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.

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

200

mouse

expressed in mouse cell line

agarose conjugate

purified immunoglobulin

primary antibodies

PT-66, monoclonal

PBS solution

immunoprecipitation (IP): suitable

IgG1

1 mg/mL binding capacity

wet ice

2-8℃

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12 - Non Combustible Liquids

WGK 3