Total & Phospho ZAP-70 Dual Detection ELISA Kit antibody 
RabMAb®
Cat.#: 6117-1
Price: $450
A Simple, Complete ELISA Kit
- All-In-One: Quick and easy sandwich ELISA Kit
- Sensitive: Useful for detection of levels of total Zap-70 and Zap-70 Phosphorylated at pY492
Description
ZAP-70, a Syk-family protein tyrosine kinase, plays a critical role in mediating T cell signal transduction in response to T cell receptor (TCR) activation (1). TCR-mediated activation of the Src-family kinases, Lck and Fyn, results in tyrosine phosphorylation of the TCR zeta and CD3 chains. These domains serve as targets for binding of ZAP-70 via its tandem SH2 domains. This binding correlates with activation of ZAP-70, a critical event in T cell activation (2). Following TCR engagement, ZAP-70 is phosphorylated on several tyrosine residues, presumably by two mechanisms: an autophosphorylation and a trans-phosphorylation by the Src-family tyrosine kinase, Lck. Lck phosphorylates Tyrosine 492 and 493 located at the activation loop of the catalytic domain of ZAP-70, leading to its activation and increased autophosphorylation (3). This assay employs the sandwich enzyme immunoassay technique that detects endogenous levels of ZAP-70 and phospho ZAP-70 pY492. Rabbit anti-ZAP-70 monoclonal antibody has been pre-coated onto six strips of a microplate. Anti-ZAP-70 pY492 monoclonal antibody has been pre-coated onto six strips of a microplate. Cell lysates are pipetted into the wells. Following extensive washing, biotinylated rabbit anti-ZAP-70 monoclonal antibody reagent is added to detect ZAP-70 and phospho ZAP-70 pY492. Following a wash to remove any unbound antibody reagent, streptavidin-HRP is added to the well. After washing away the unbound streptavidin-HRP, TMB substrate is added to the wells to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of ZAP-70 protein and phospho ZAP-70 pY492.
Recommended Applications
ELISA
Products Data
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| Fig 1. Main steps of the ZAP-70 Total and Phospho Sandwich ELISA assay. |
Components
- Rabbit ZAP-70 Monoclonal Anitbody Coated Microwells (48 microwells)
- Rabbit ZAP-70 Phospho (pY492) Monoclonal Antibody Coated Microwells (48 microwells)
- Biotinylated ZAP-70 Total Antibody Reagent (96 reactions, 11mLs)
- Streptavidin-HRP Reagent
- Antigen/Antibody Diluent Buffer
- ELISA Washing Buffer
- TMB A Substrate Solution
- TMB B Substrate Solution
- Stop Solution
Storage Condition and Buffer
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