Analysis Note

ControlSuggested Positive Antigen Control: Cat. # 12-302, EGF-stimulated A431 cell lysate is provided as a free positive antigen control for western immunoblotting. Aliquot as desired, refreeze immediately, and store at -20°C. The lysate is stable for 6 months at -20°C. Before use, add 2.5 µL of 2-mercaptoethanol per 100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for immunoblot analysis.

Application

Research CategorySignaling

4G10 Platinum, Anti-Phosphotyrosine (mouse monoclonal cocktail IgG2b) detects levels of Phosphotyrosine proteins & has been published & validated for use in WB, ICC, ELISA, IHC, IF, Flow & IP.

Research Sub CategoryGeneral Post-translation Modification

Immunoprecipitation: 2-4 µL of a previous lot can immunoprecipitate quantitatively the phosphotyrosine containing proteins in the lysate of a confluent culture (10cm dish) of cells expressing an activated tyrosine kinase. To preserve phosphotyrosine, add 0.2 mM sodium orthovanadate to the lysis buffer.

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

The development of the anti-phosphotyrosine, clone 4G10™ in 1989 was a monumental discovery for researchers. 4G10™ was the first and is the best single monoclonal antibody for the detection tyrosine phosphorylation. 4G10™ is well known for its sensitivity and its ability to detect multiple tyrosine phosphorylations on numerous substrates. It has been validated by thousands of scientific and medical researchers in virtually every application and tyrosine target over the past 2 decades. To improve on something that hundred have tried and no one has succeeded, we pooled 4G10™ with the next most highly regarded anti-phosphotyrosine, clone PY20 to make 4G10™ Platinum. PY20 itself is a very poor substitute for 4G10™, but its additive effect allow for a greater level of detection on more substrates that even 4G10™ alone was not capable of.

Immunogen

4G10™ used Phosphotyramine coupled to KLH and PY20 used phosphotyrosine conjugated to carrier protein

Legal Information

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

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

Physical form

Protein G-Sepharose™ Chromatography

Format: Purified

Quality

Evaluated by Western Blot on a modified RIPA lysate from EGF-treated human A431 carcinoma cells.

Western Blot Analysis: 1:1,000-1:2,000 of this antibody detected tyrosinephosphorylated proteins in a modified RIPA lysate from EGF-treated human A431 carcinoma cells.

Specificity

Tyrosine-phosphorylated proteins from all species

Storage and Stability

Stable for 1 year at from date of receipt.

Target description

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