ZAP-70 Phospho (pY492) ELISA Kit antibody 
RabMAb®
Cat.#: 6113-1
Price: $450
A Simple, Complete ELISA Kit
- All-In-One: Quick and easy sandwich ELISA Kit
- Sensitive: Useful for detection of levels of 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 (3). This assay employs the sandwich enzyme immunoassay technique that detects endogenous levels of ZAP-70 when phosphorylated at Tyrosine 492 (Fig 1). Rabbit anti-ZAP-70 monoclonal antibody has been pre-coated onto a microplate (Step 1). Cell lysate is pipetted into the wells (Step 2). Following extensive washing, biotinylated rabbit anti-ZAP- 70 Phospho (pY492) monoclonal antibody reagent is added (Step 3). Following a wash to remove any unbound antibody reagent, streptavidin- HRP is added to the well (Step 4). After washing away the unbound streptavidin-HRP, a substrate solution is added to the wells to develop color (Step 5). The magnitude of the absorbance for this developed color is proportional to the amount of phosphorylated ZAP-70.
Recommended Applications
ELISA
Products Data
 |  |
| Fig. 1 Main steps of the ZAP-70 Phospho (pY492) ELISA Kit |
Components
- Dry Plate with stripwells (96)
- Biotinylated ZAP-70 Phospho (pY492) Antibody Reagent
- Streptavidin-HRP Reagent
- Antigen/Antibody Diluent Buffer
- ELISA Washing Buffer
- TMB A Substrate Solution
- TMB B Substrate Solution
- Stop Solution
Storage Condition and Buffer
Store at -20 °C. Buffer: 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol, 0.01% sodium azide and 0.05% BSA. Stable for 12 months from date of receipt.Related Products
| Product Name | Catalog # |
|---|---|
| Goat Anti-Rabbit IgG (H+L) HRP | 3053-1 |
| Goat Anti-Rabbit IgG polymerized HRP | 3051-1 |
| Cell Lysis Buffer | 3500-1 |
| Goat Anti-Rabbit IgG HRP Antibody | 3056-1 |
| Goat Anti-Rabbit IgG DyLight® 405 | 3063-1 |
| Goat Anti-Rabbit IgG DyLight® 488 | 3064-1 |
| Goat Anti-Rabbit IgG DyLight® 550 | 3065-1 |
| Goat Anti-Rabbit IgG DyLight® 594 | 3066-1 |
| Goat Anti-Rabbit IgG DyLight® 633 | 3067-1 |
| Goat Anti-Rabbit IgG DyLight® 650 | 3068-1 |
