Extracts of NIH3T3 cells stably expressing c-Src Y527F mutant protein were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was left untreated (1-4) or treated with lambda (λ) phosphatase (5), blocked with a 5% BSA-TBST buffer overnight at 4°C, then incubated with the cortactin [pY466] (mouse) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphotyrosine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase (Prod # ALI4405) and signals were detected using the Tropix WesternStar™ method.
|Tested species reactivity||Mouse|
|Published species reactivity||All, Tag, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||antiserum was produced against a chemically synthesized phosphopeptide derived from the region of mouse cortactin that contains tyrosine 466 (tyrosine 470 in the human sequence).|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Western Blot (WB)||0.1-1.0 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
Cortactin is an 80/85 kDa cytoskeleton protein that facilitates assembly of cortical actin. Cortactin is widely expressed in most adherent cells and is a prominent substrate of protein-tyrosine kinase Src in vivo and in vitro. The protein sequence of cortactin contains multiple amino-terminal tandem repeats of a unique 37-amino acid sequence, which associates with actin, and a Src homology 3 (SH3) domain at the carboxyl terminus. Between the repeat and the SH3 domain, there is an alpha-helical structure followed by a proline-rich region.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Cortactin tyrosine phosphorylation requires Rac1 activity and association with the cortical actin cytoskeleton.||Head JA,Jiang D,Li M,Zorn LJ,Schaefer EM,Parsons JT,Weed SA||Molecular biology of the cell (14:3216)||2003|