| Product Includes | Product # | Quantity | Color |
|---|---|---|---|
| FastScan™ ELISA Microwell Strip Plate, 96 Well | 53257 | 96 tests | |
| Zap-70 Rabbit Capture mAb | 43665 | 1 ea | Green (Lyophilized) |
| Phospho-Zap-70 (Tyr319) Rabbit HRP-linked mAb | 30808 | 1 ea | Red (Lyophilized) |
| FastScan™ ELISA Capture Antibody Diluent | 16076 | 3 ml | Green |
| FastScan™ ELISA HRP Antibody Diluent | 28120 | 3 ml | |
| TMB Substrate | 7004 | 11 ml | |
| STOP Solution | 7002 | 11 ml | |
| Sealing Tape | 54503 | 1 ea | |
| ELISA Wash Buffer (20X) | 9801 | 25 ml | |
| FastScan™ ELISA Cell Extraction Buffer (5X) | 69905 | 10 ml | |
| FastScan™ ELISA Cell Extraction Enhancer Solution (50X) | 25243 | 1 ml | |
| FastScan™ ELISA Kit #47223 Positive Control Type 2 | 68594 | 1 ea |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The FastScan™ Phospho-Zap-70 (Tyr319) ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Zap-70 when phosphorylated at Tyr319. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with phospho-Zap-70 (Tyr319) in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of phospho-Zap-70 (Tyr319).
*Antibodies in kit are custom formulations specific to kit.
IMPORTANT: This FastScan™ ELISA kit requires 4 washes at Step 6 of the protocol.
Specificity/Sensitivity
Background
The Syk family protein tyrosine kinase Zap-70 is expressed in T and NK cells and plays a critical role in mediating T cell activation in response to T cell receptor (TCR) engagement (1). Following TCR engagement, Zap-70 is rapidly phosphorylated on several tyrosine residues through autophosphorylation and transphosphorylation by the Src family tyrosine kinase Lck (2-6). Tyrosine phosphorylation correlates with increased Zap-70 kinase activity and downstream signaling events. Expression of Zap-70 is correlated with disease progression and survival in patients with chronic lymphocytic leukemia (7,8).
- Chu, D.H. et al. (1998) Immunol Rev 165, 167-80.
- Iwashima, M. et al. (1994) Science 263, 1136-9.
- Neumeister, E.N. et al. (1995) Mol Cell Biol 15, 3171-8.
- Chan, A.C. et al. (1995) EMBO J 14, 2499-508.
- Williams, B.L. et al. (1999) EMBO J 18, 1832-44.
- Di Bartolo, V. et al. (1999) J Biol Chem 274, 6285-94.
- Wiestner, A. et al. (2003) Blood 101, 4944-51.
- Crespo, M. et al. (2003) N Engl J Med 348, 1764-75.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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