Analysis Note

ControlPervanadate-treated human A431 cell extracts, EGF-treated human A431 cells.

Application

For use in applications in which an HRP conjugate is advantageous, such as WB and IP.

Research CategorySignaling

Research Sub CategoryGeneral Post-translation Modification

Anti-Phosphotyrosine Antibody, clone 4G10™, HRP conjugate detects level of Phosphotyrosine & has been published & validated for use in ELISA & WB.

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.

General description

Anti-Phosphotyrosine antibody (05-321), cross-linked to horseradish peroxidase

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
IgG2bκ mouse monoclonal antibody produced in vitro by mouse-mouse hybridoma 4G10™. Purified by Protein G-Sepharose chromatography and cross-linked to horseradish peroxidase.

Immunogen

Phosphotyramine-KLH.

Legal Information

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

Kathon is a registered trademark of Rohm and Haas Co.

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

Physical form

200 µg of IgG2bκ-HRP conjugate in 100 µL PBS containing 0.05% Kathon^® . Sufficient for 10 western blots, if used at the optimum concentration of 2 µg/mL in 10 mL blocking buffer. Liquid.

Protein G-Sepharose™ Chromatography

Quality

Routinely evaluated in western blot on RIPA lysates from EGF-treated human A431 carcinoma cells.

Western Blot Analysis:
Tested at 0.5-2 µg/mL, with optimal performance at 2 µg/mL (10 µL per 10 mL blocking buffer). This lot detected tyrosine-phosphorylated proteins in RIPA lysates from EGF-treated human A431 carcinoma cells.

Specificity

Recognizes tyrosine-phosphorylated proteins from all species.

Storage and Stability

Stable for 6 months at 2-8°C from date of receipt. NOTE: DO NOT FREEZE. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
NOTE: For optimal performance, remove HRP precipitate by centrifuging vial prior to use.

Target description

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