Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

Zap-70 Blocking Peptide #1160

Description

This peptide can be used to block Zap-70 (99F2) Rabbit mAb #2705.

Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Zap-70 (99F2) Rabbit mAb #2705 signal completely in both immunohistochemistry and Western blotting.

Western Blotting

Western Blotting

Western Blot analysis of extracts from Jurkat and Ramos cells, using Zap-70 (99F2) Rabbit mAb #2705 (left) or the same antibody preincubated with Zap-70 Blocking Peptide (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical/immunofluorescent analysis of paraffin-embedded human tonsil, using Zap-70 (99F2) Rabbit mAb #2705 (left, green), or the same antibody preincubated with Zap-70 Blocking Peptide (right). Nuclei were counterstained using DAPI.

Applications

Use as a blocking reagent to evaluate the specificity of antibody reactivity in Western immunoblotting and immunohistochemistry protocols.

Directions for Use

For immunohistochemistry, add 2 µl of antibody #2705 and 2 µl of blocking peptide to 0.096 ml of buffer, and incubate at least 30 minutes before allowing to react with the slides. For Western immunoblotting, add 10 µl of antibody and 10 µl of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room temperature for 2 hours before allowing to react with the blot.

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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