Order 3, Get Your 4th Free!* | Learn More >>
7528
HTScan® CDK3/CycE Kinase Assay Kit
Cellular Assay Kits
Assay Kit

HTScan® CDK3/CycE Kinase Assay Kit #7528

Citations (0)
HTScan® CDK3/CycE Kinase Assay Kit: Image 1

Figure 2. Dose dependence curve of CDK3/CycE kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK3/CycE kinase. In a 50 µl reaction, increasing amounts of CDK3/CycE 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® CDK3/CycE Kinase Assay Kit: Image 2

Figure 5. Staurosporine inhibition of CDK3/CycE kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK3/CycE kinase. In a 50 µl reaction, 50 ng CDK3/CycE, 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® CDK3/CycE Kinase Assay Kit: Image 3

Figure 3. Peptide concentration dependence of CDK3/CycE kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK3/CycE kinase. In a 50 µl reaction, 50 ng of CDK3/CycE 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® CDK3/CycE Kinase Assay Kit: Image 4

Figure 1. Time course of CDK3/CycE kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK3/CycE kinase. In a 50 µl reaction, 50 ng CDK3/CycE and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

HTScan® CDK3/CycE Kinase Assay Kit: Image 5

Figure 1. CDK3/CycE kinase activity was measured in a radioisotopic filter binding assay using the following reaction conditions: 60 mM HEPES-NaOH, pH 7.5, 3 mM MgCl2, 3 mM MnCl2, 3 µM Na-orthovanadate, 1.2 mM DTT, ATP (variable), 10 ng/50 µl PEG20,000, Substrate: Rb CTF, 5 µg /50 µl, recombinant CDK3/CycE: 2 Units/50 µl.

To Purchase # 7528

Product Description

The kit provides a means of performing kinase activity assays with recombinant human CDK3/CycE kinase. It includes active CDK3/CycE 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-Rb (Ser807/811): GST-CDK3 kinase: 68 kDa, GST-CycE: 72 kDa.

Background

Cyclins and cyclin-dependent kinases (CDK) are key regulators in mammalian cell cycle. Regulation of these complexes occurs through cyclin production and its destruction, relocation, inhibitory/activating phoshorylation, relocation and modification by other proteins. Each cyclin associates with one or two CDKs and most CDKs associate with one or two cyclins (1-3). CDK1 forms a complex with cyclin A/B and regulates phosphorylation of cytoskeleton proteins involved in mitosis. CDK2 and CDK3 form complexes with cyclin E, which regulate the G1-S phase transition while the CDK2/CycA complex regulates S phase progression (4,5). CDK4/CycD and CDK6/CycD are activated by mitogenic signaling during early G1 and progressively accumulate as cells transition through this phase of the cell cycle. CDK5 is activated in postmitotic neurons and regulates neuron migration during brain development (6). CDK7/CycH is believed to form a link between transcription and cell cycle. CDK8/CycC and CDK9/CycT are involved in transcription (1,2). The kinase activity of CDKs is tightly regulated by phosphorylation and protein-protein interactions. Activation of CDKs requires binding to a specific cyclin and phosphorylation of a conserved threonine residue in a region called the T loop. Examining the phosphorylation of peptides by CDK/cyclin complexes suggests that both CDKs and cyclins play a role in recognizing substrates. A consensus sequence, (S/T)PX(R/K), is identified in the peptides that are phosphorylated by CDK/cyclins.

  1. Schang, L.M. (2002) J. Antimicrob. Chemother. 50, 779-792.
  2. Murray, A.W. (2004) Cell 116, 221-234.
  3. Chow, J.P. et al. (2003) J. Biol. Chem. 278, 40815-40828.
  4. Hofmann, F. and Livingston, D.M. (1996) Genes Dev. 10, 851-861.
  5. Golsteyn, R.M. (2005) Cancer Lett. 217, 129-138.
  6. Xie, Y. and Tsai, L.H. (2004) Cell Cycle 3, 108-110.
  7. Holmes, J.K. and Solomon, M.J. (1996) J Biol Chem 271, 25240-6.

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.