Order 3, Get Your 4th Free!* | Learn More >>
7779
HTScan® Syk Kinase Assay Kit
Cellular Assay Kits
Assay Kit

HTScan® Syk Kinase Assay Kit #7779

Citations (0)
HTScan® Syk Kinase Assay Kit: Image 1

Figure 3. Dose dependence curve of Syk kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1310) by Syk kinase. In a 50 µl reaction, increasing amounts of Syk and 1.5 µM substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

HTScan® Syk Kinase Assay Kit: Image 2

Figure 5. Staurosporine inhibition of Syk kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of Syk substrate peptide (#1310) by Syk kinase. In a 50 µl reaction, 50 ng Syk, 1.5 µM substrate peptide, 20 µM ATP and increasing amounts of staurosporine were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

HTScan® Syk Kinase Assay Kit: Image 3

Figure 4. Peptide concentration dependence of Syk kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1310) by Syk kinase. In a 50 µl reaction, 50 ng of Syk and increasing concentrations of substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

HTScan® Syk Kinase Assay Kit: Image 4

Figure 2. Time course of Syk kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of Syk substrate peptide (#1310) by Syk kinase. In a 50 µl reaction, 50 ng Syk and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

HTScan® Syk Kinase Assay Kit: Image 5

Figure 1. Syk kinase activity was measured in a radiometric assay using the following reaction conditions: 5 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 0.4 mM EDTA, 4 mM MgCl2, 2.5 mM MnCl2, 0.05 mM DTT, 50 μM ATP, Substrate: Poly EY, 400 ng/μL, and recombinant Syk: variable.

To Purchase # 7779

Product Description

The kit provides a means of performing kinase activity assays with recombinant human Syk kinase. It includes active Syk kinase (supplied as a GST fusion protein), a biotinylated peptide substrate and a phospho-tyrosine antibody for detection of the phosphorylated form of the substrate peptide.

Molecular Formula

Peptide substrate, Biotin-Gastrin Precursor (Tyr87): 2,853 Daltons. GST-Syk Kinase: 104 kDa

Background

Syk is a protein tyrosine kinase that plays an important role in intracellular signal transduction in hematopoietic cells (1-3). Syk interacts with immunoreceptor tyrosine-based activation motifs (ITAMs) located in the cytoplasmic domains of immune receptors (4). It couples the activated immunoreceptors to downstream signaling events that mediate diverse cellular responses, including proliferation, differentiation, and phagocytosis (4). There is also evidence of a role for Syk in nonimmune cells and investigators have indicated that Syk is a potential tumor suppressor in human breast carcinomas (5). Tyr323 is a negative regulatory phosphorylation site within the SH2-kinase linker region in Syk. Phosphorylation at Tyr323 provides a direct binding site for the TKB domain of Cbl (6,7). Tyr352 of Syk is involved in the association of PLCγ1 (8). Tyr525 and Tyr526 are located in the activation loop of the Syk kinase domain; phosphorylation at Tyr525/526 of human Syk (equivalent to Tyr519/520 of mouse Syk) is essential for Syk function (9).

  1. Cheng, A.M. and Chan, A.C. (1997) Curr Opin Immunol 9, 528-33.
  2. Kurosaki, T. (1997) Curr Opin Immunol 9, 309-18.
  3. Chu, D.H. et al. (1998) Immunol Rev 165, 167-80.
  4. Turner, M. et al. (2000) Immunol Today 21, 148-54.
  5. Coopman, P.J. et al. (2000) Nature 406, 742-7.
  6. Deckert, M. et al. (1998) J Biol Chem 273, 8867-74.
  7. Rao, N. et al. (2001) EMBO J 20, 7085-95.
  8. Law, C.L. et al. (1996) Mol Cell Biol 16, 1305-15.
  9. Zhang, J. et al. (2000) J Biol Chem 275, 35442-7.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
DELFIA is a registered trademark of PerkinElmer, Inc.