Analysis Note

Positive ControlVanadate-treated A431 cells or 8-pCPT-treated platelets

Application

ELISA (0.05 µg/ml)r>Immunoblotting (0.1 µg/ml, chemiluminescence; 1 µg/ml, colorimetric)r>Immunocytochemistry (1-10 µg/ml)r>Immunoprecipitation (1-10 µg/10⁶ vanadate-treated A431 cells)

General description

Recognizes the ~46-50 kDa VASP protein phosphorylated at Ser²³⁹.

This PhosphoDetect™ Anti-VASP (pSer²³⁹) Mouse mAb (16C2) is validated for use in ELISA, Immunoblotting, ICC, IP for the detection of VASP (pSer²³⁹).

Mouse monoclonal antibody purified from serum-free cell culture supernatant by thiophilic adsorption and size exclusion chromatography. Recognizes the ~46-50 kDa VASP protein phosphorylated at Ser²³⁹.

Immunogen

a synthetic phosphopeptide corresponding to amino acids surrounding the Ser²³⁹ phosphorylation site of human VASP, conjugated to KLH

Human

Legal Information

Triton is a trademark of The Dow Chemical Company or an affiliated company of Dow

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

Other Notes

Reinhard, M., et al. 1999. In: Guidebook to the Cytoskeletal and Motor Proteins (Eds. Kreis, T., and Vale, R.), Oxford University Press, Oxford, 168.Schwarz, U.R., et al. 1999. Thromb. Haemost.82, 1145.Smolenski, A., et al. 1998. J. Biol. Chem.273, 20029.

Does not recognize the non-phosphorylated form of VASP. Suitable for monitoring the phosphorylation state of VASP Ser²³⁹ as well as PKA/PKG activity. For immunocytochemistry, SNP or 8-pCPT-cGMP treated platelets are fixed with 4% paraformaldehyde in PBS for 15 min. on ice, then permeabilized with 0.2% Triton^™ X-100 detergent in PBS for 10 min at room temperature. For immunoblotting, block with 1% hemoglobin in PBS, 0.3% Triton™ X-100 detergent, and 0.05% Tween™-20 detergent. Variables associated with assay conditions will dictate the proper working dilution.

Packaging

100 µg in Plastic ampoule

Physical form

Lyophilized from 2X PBS, PEG, sucrose.

Reconstitution

Reconstitute with 1 ml of dH₂O. Following reconstitution, refrigerate (4°C) for short-term storage or aliquot, flash-freeze in liquid nitrogen, and freeze (-70°C) for long-term storage. Stock solutions are stable for up to 3 months at 4°C or for up to 1 year at -70°C. Thaw aliquots at 37°C. Avoid freeze/thaw cycles of solutions.

Warning

Toxicity: Standard Handling (A)