Interested in promotions? | Click here >>
7508
HTScan® ROCK2 Kinase Assay Kit
Cellular Assay Kits
Assay Kit

HTScan® ROCK2 Kinase Assay Kit #7508

Citations (0)
HTScan® ROCK2 Kinase Assay Kit: Image 1

Figure 3. Dose dependence curve of ROCK2 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of substrate peptide (#1300) by ROCK2 kinase. In a 50 µl reaction, increasing amounts of ROCK2 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® ROCK2 Kinase Assay Kit: Image 2

Figure 5. Staurosporine inhibition of ROCK2 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of ROCK2 substrate peptide (#1300) by ROCK2 kinase. In a 50 µl reaction, 50 ng ROCK2, 1.5 µM substrate peptide, 50 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® ROCK2 Kinase Assay Kit: Image 3

Figure 4. Peptide concentration dependence of ROCK2 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of substrate peptide (#1300) by ROCK2 kinase. In a 50 µl reaction, 50 ng of ROCK2 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® ROCK2 Kinase Assay Kit: Image 4

Figure 2. Time course of ROCK2 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibodty #5070 to detect phosphorylation of ROCK2 substrate peptide (#1300) by ROCK2 kinase. In a 50 µl reaction, 50 ng ROCK2 and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

HTScan® ROCK2 Kinase Assay Kit: Image 5

Figure 1. ROCK2 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, 5 mM MgCl2, 0.05 mM DTT, 50 μM ATP, Substrate: S6K substrate peptide (KRRRLASLR) 400 ng/μL, and variable amounts of recombinant ROCK2.

Inquiry Info.# 7508

Product Description

The kit provides a means of performing kinase activity assays with recombinant human ROCK2 kinase. It includes active ROCK2 kinase (supplied as a GST fusion protein), a biotinylated peptide substrate and a phospho-serine/threonine antibody for detection of the phosphorylated form of the substrate peptide.

Molecular Formula

Peptide substrate, Biotin-PLK (Ser137): 1,945 Daltons. GST-ROCK2 Kinase: 88 kDa.

Background

ROCK (Rho-associated kinase), a family of serine/threonine kinases, is an important downstream target of Rho-GTPase and plays an important role in Rho-mediated signaling. Two isoforms of ROCK have been identified: ROCK1 and ROCK2. ROCK is composed of N-terminal catalytic, coiled-coil, and C-terminal PH (pleckstrin homology) domains. The C-terminus of ROCK negatively regulates its kinase activity (1,2). Caspase-3-induced cleavage of ROCK1 and direct cleavage of ROCK2 by granzyme B (grB) activates ROCK and leads to phosphorylation of myosin light chain and inhibition of myosin phosphatase (3). This phosphorylation may account for the mechanism by which Rho regulates cytokinesis, cell motility, cell membrane blebbing during apoptosis, and smooth muscle contraction (4-6).

  1. Nakagawa, O. et al. (1996) FEBS Lett. 392, 189-193.
  2. Lee, J.H. et al. (2004) J. Cell. Biol. 167, 327-337.
  3. Sebbagh, M. et al. (2005) J. Exp. Med. 201, 465-471.
  4. Amano, M. et al. (1996) J. Biol. Chem. 271, 20246-20249.
  5. Kureishi, Y. et al. (1997) J. Biol. Chem. 272, 12257-12260.
  6. Totsukawa, G. et al. (2000) J. Cell Biol. 150, 797-806.

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.