Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total Zap-70 Sandwich ELISA Antibody Pair #7850

Kit Includes Volume Cap Color
Zap-70 Capture Ab (100X) 0.4 ml Pink
Zap-70 Detection Ab (100X) 0.4 ml Blue
Anti-rabbit IgG, HRP-linked Antibody (1000X) 0.04 ml Red

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Important Ordering Details

Product is assembled upon order to ensure maximum activity. Domestic: Please allow up to two weeks for your order to be processed and shipped. International: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.

Protocols

Description

CST's PathScan® Total Zap-70 Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total Zap-70 Sandwich ELISA Kit #7172. Capture and detection antibodies (100X stocks) and HRP-linked secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Zap-70 capture antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a Zap-70 detection antibody and anti-Rabbit IgG, HRP-linked antibody. HRP substrate TMB is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of total Zap-70 protein.

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

The relationship between protein concentration of lysates from untreated and H2O2-treated Jurkat cells and the absorbance at 450 nm using PathScan® Total Zap-70 Sandwich ELISA Antibody Pair #7850 is shown. Jurkat cells (0.8 x 106 cells/ml) were treated with H2O2 (2 mM) for 2 min at 25ºC 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, 167-180.
  2. Iwashima, M. et al. (1994) Science 263, 1136-1139.
  3. Neumeister, E.N. et al. (1995) Mol. Cell Biol. 15, 3171-3178.
  4. Chan, A.C. et al. (1995) EMBO J. 14, 2499-2508.
  5. Williams, B.L. et al. (1999) EMBO J. 18, 1832-1844.
  6. Di Bartolo, V. et al. (1999) J. Biol. Chem. 274, 6285-6294.
  7. Wiestner, A. et al. (2003) Blood 101, 4944-4951.
  8. Crespo, M. et al. (2003) N. Engl. J. Med. 348, 1764-1775.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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