New England Biolabs Canada
 

Product Pathways -

FastScan™ Total Zap-70 ELISA Kit #23925

Item# Description List Price Web Price Qty
23925C FastScan™ Total Zap-70 ELISA Kit - 1 Kit $699.00
$629.10
ADD TO CART
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS

Protocols

Description

The FastScan™ Total Zap-70 ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Zap-70. 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 Zap-70 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 Zap-70. Antibodies in kit are custom formulations specific to kit.

Cell-based assay

Cell-based assay

Figure 1. Treatment of Jurkat cells with H2O2 stimulates phosphorylation of Zap-70 at Tyr319 but does not affect the level of total Zap-70. The relationship between lysate protein concentration from untreated and H2O2-treated Jurkat cells and the absorbance at 450 nm using the FastScan™ Total Zap-70 ELISA Kit #23925 is shown in the upper figure. The corresponding western blots using Zap-70 antibody (left panel) and phospho-Zap-70 (Tyr319) antibody (right panel) are shown in the lower figure. After serum starvation, Jurkat cells were treated with 11 mM H2O2 for 3 minutes at 37°C and then lysed.

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).

  1. Chu, D.H. et al. (1998) Immunol Rev 165, 167-80.
  2. Iwashima, M. et al. (1994) Science 263, 1136-9.
  3. Neumeister, E.N. et al. (1995) Mol Cell Biol 15, 3171-8.
  4. Chan, A.C. et al. (1995) EMBO J 14, 2499-508.
  5. Williams, B.L. et al. (1999) EMBO J 18, 1832-44.
  6. Di Bartolo, V. et al. (1999) J Biol Chem 274, 6285-94.
  7. Wiestner, A. et al. (2003) Blood 101, 4944-51.
  8. Crespo, M. et al. (2003) N Engl J Med 348, 1764-75.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


 

Toll Free: 1-800-387-1095

Fax: 1-800-563-3789

info.ca@neb.com

orders.ca@neb.com

About NEBTerms of Sale Web Discount Shipping Information Web Site Disclaimer Privacy Policy NEB USA Cell Signaling Technology Sitemap
Contents ¬©New England Biolabs Ltd.  New England Biolabs Ltd. is the exclusive Canadian distributor for Cell Signaling Technology, Inc.  New England Biolabs, Inc. is an ISO 9001 certified company.

Search another product:

 

Item has been added to the cart

 

Item has been added to the favourites