Anti-Phosphotyrosine Antibody, clone 4G10^®, Biotin Conjugate

Anti-phosphotyrosine monoclonal antibody, clone 4G10 (cat.# 05-321), cross-linked to biotin.

Dependent upon the molecular weight of the tyrosine phosphorylated protein being detected.

Some of the tyrosine residues can be tagged with a phosphate group (phosphorylated) by protein kinases. (In its phosphorylated state, it is referred to as phosphotyrosine.). Tyrosine phosphorylation is considered as one of the key steps in signal transduction and regulation of enzymatic activity. The advent of anti-phospho-tyrosine antibodies is one of significant events in signal transduction research. Before the availability of anti-phosphotyrosine antibodies, tyrosyl phospyhorylation of proteins and enzymes was investigated through hazardous and time-consuming radioactive experiments. Anti-phosphotyrosine antibodies are commonly used in western blots after the targeted proteins have been immunoprecipitated to measure the tyrosyl phosphorylation of the proteins. Anti-phosphotyrosine antibodies are also directly used on cell lysate to examine the overall change of tyrosine phosphorylation level in response to various treatments.

Description
Anti-phosphotyrosine monoclonal antibody, clone 4G10 (cat. # 05-321), cross-linked to biotin.

Phosphotyramine-KLH.

Anti-Phosphotyrosine Antibody, clone 4G10, Biotin Conjugate detects tyrosine phosphorylated proteins in all species. This unique monoclonal antibody is published in peer reviewed journals and validated for use in IC, IP, WB.

Immunoprecipitation:
4 μg of a previous lot, used in conjunction with Streptavidin, agarose conjugate (Catalog # 16-126), immunoprecipitated phosphotyrosine containing proteins from a lysate of EGFstimulated A431 cells.
Note: To preserve phosphotyrosine, add 0.2 mM sodium orthovanadate to the lysis buffer.

Immunocytochemistry:
5-10 μg/mL of a previous lot gave positive immunostaining of EGF-stimulated A431 cells that had been fixed with ethanol:acetic acid [1:1].

Application Notes
For use in applications in which a biotin conjugate is advantageous, such as WB and IC.

Research Category
Signaling

Research Sub Category
General Post-translation Modification

Expected to cross-react with rat and mouse.

Recognizes tyrosine-phosphorylated proteins from all species.

PBS containing 0.05% sodium azide.

Protein G Purified

9 months at 2-8°C from date of shipment. NOTE: DO NOT FREEZE. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

Control
Pervanadate-treated human A431 cell extracts or EGF-treated human A431 cells

Routinely evaluated by Western Blot on EGF treated A431 lysates.

Western Blot Analysis:
1:500 dilution of this lot detected Tyrosine phosphorylated proteins on 10 μg of EGF treated A431 lysates.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

4G10 is a registered trademark of Upstate Group, Inc.

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

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.