Analysis Note

ControlChick embryo fibroblast (CEF) cells expressing human wild-type Src protein

Application

Research CategorySignaling

Detect phospho-Src (Tyr416) using this Anti-phospho-Src (Tyr416) Antibody, clone 9A6 validated for use in WB & IP.

Research Sub CategoryCytoskeletal Signaling

Immunoprecipitation/Immunoblot Analysis: 4 μg/mL of a previous lot detected phospho-Src immunoprecipitated from 3T3 cell lysates with 10 μg of anti-Src (Catalog # 05-184), but not after phosphatase treatment with YOP.

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

Src (also known as pp60src) is a non receptor Tyrosine Kinase involved in signal transduction in many biological systems and implicated in the development of human tumors. There are two critical phosphorylation sites of tyrosine on Src, tyrosine 418 and tyrosine 529 (referring to human Src sequence). The tyrosine 418 is located in the catalytic domain and is one of the autophosphorylation sites. Full catalytic activity of Src requires phosphorylation of tyrosine 418. This region of Src is also highly conserved in all of the related Src-family kinases, and thus prior immunoprecipitation may be required to identify which Src family member is being activated.

Specific src isoform is unknown

Immunogen

KLH-conjugated, synthetic peptide containing phosphotyrosine in the sequence context corresponding to tyrosine 416 of avian Src. Clone 9A6.

Legal Information

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

Other Notes

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

Physical form

Purified mouse monoclonal IgG3 lyophilized from 1 mL 2x PBS / 0.09 % Na-azide / PEG and Sucrose. Reconstitute with 1 mL H2O (15 min, RT). Lyophilized. Store at -20°C.

Thiophilic adsoption and size exclusion chromatography

Format: Purified

Quality

Evaluated by immunoprecipitating Src from 3T3 cell lysates with anti-Src (Catalog #05-184), but not after phosphatase treatment with YOP.

Src Autophosphorylation/Western Blotting Analysis: 0.5-2 μg/mL of this antibody strongly detected recombinant Src (Catalog # 14-326) after incubation with Mg/Mn/ATP. Phospho-specificity was confirmed by dephosphorylation of the Src protein with λ-phosphatase.

Specificity

Recognizes phospho-Src, Mr 60kDa. The antibody does not crossreact with the nonphosphorylated form of Src or with unrelated phosphorylation sites. Predicted to recognize all Src family members phosphorylated at the tyrosine corresponding to Tyr416 of avian Src.

Storage and Stability

Stable for 1 year at from date of receipt.

Target description

60 kDa