Interested in promotions? | Click here >>
7639
HTScan® Mst4 Kinase Assay Kit
Cellular Assay Kits
Assay Kit

HTScan® Mst4 Kinase Assay Kit #7639

Citations (0)
HTScan® Mst4 Kinase Assay Kit: Image 1

Figure 3. Dose dependence curve of Mst4 kinase activity: DELFIA® data generated using Phospho-Ezrin (Thr567)/Radixin (Thr564)/Moesin (Thr558) Antibody #3141 to detect phosphorylation of substrate peptide (#1344) by Mst4 kinase. In a 50 µl reaction, increasing amounts of Mst4 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® Mst4 Kinase Assay Kit: Image 2

Figure 5. Staurosporine inhibition of Mst4 kinase activity: DELFIA® data generated using Phospho-Ezrin (Thr567)/Radixin (Thr564)/Moesin (Thr558) Antibody #3141 to detect phosphorylation of Mst4 substrate peptide (#1344) by Mst4 kinase. In a 50 µl reaction, 50 ng Mst4, 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® Mst4 Kinase Assay Kit: Image 3

Figure 4. Peptide concentration dependence of Mst4 kinase activity: DELFIA® data generated using Phospho-Ezrin (Thr567)/Radixin (Thr564)/Moesin (Thr558) Antibody #3141 to detect phosphorylation of substrate peptide (#1344) by Mst4 kinase. In a 50 µl reaction, 50 ng of Mst4 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® Mst4 Kinase Assay Kit: Image 4

Figure 2. Time course of Mst4 kinase activity: DELFIA® data generated using Phospho-Ezrin (Thr567)/Radixin (Thr564)/Moesin (Thr558) Antibody #3141 to detect phosphorylation of Mst4 substrate peptide (#1344) by Mst4 kinase. In a 50 µl reaction, 50 ng Mst4 and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

HTScan® Mst4 Kinase Assay Kit: Image 5

Figure 1. Mst4 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), 2.5 µg/50 µl PEG20.000, Substrate: Myelin basic protein, 5 µg/50 µl, recombinant Mst4: 100 ng/50 µl.

To Purchase # 7639

Product Description

The kit provides a means of performing kinase activity assays with recombinant active human Mst4 kinase. It includes active Mst4 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, Ezrin (Thr567)/Radixin (Thr564)/Moesin (Thr558) Biotinylated Peptide #1344: 1,958 Daltons, GST-Mst4 Kinase: 76 kDa.

Background

MST kinases, members of the STE20 family of kinases, are upstream activators of MAPK pathways that regulate processes such as apoptosis, morphogenesis and cytoskeletal rearrangements. The amino-terminal kinase domains of MST kinases are highly similar to the kinase domains of yeast STE20 kinase and other p21-activated kinases (1). The carboxy-terminal regions of MST1 and MST2 contain dimerization and inhibitory domains (1-3). Dimerization and phosphorylation at the activation loop results in translocation of MST1 from the cytosol to the nucleus (3). Growing evidence indicates that MST1, MST2 and MST3 are activated by apoptotic signals as well as other stress conditions (4-6). Complete activation of MST1 requires both phosphorylation and caspase-mediated cleavage (4). Sequence alignment of the activation loop of the GCK family indicates that Thr183 of MST1 and Thr180 of MST2 are the conserved residues and are critical for activation of the kinases. Activated MST kinases may rely on p38 MAPK and JNK pathways to amplify apoptotic signals (5). Phosphorylation at Ser327 of MST1, which is close to the caspase-3 recognition site, inhibits caspase-mediated cleavage (4).

  1. Dan, I. et al. (2001) Trends Cell Biol 11, 220-30.
  2. Creasy, C.L. et al. (1996) J Biol Chem 271, 21049-53.
  3. Lee KK and Yonehara S (2002) J Biol Chem 277, 12351–8
  4. Graves, J.D. et al. (2001) J Biol Chem 276, 14909-15.
  5. Lee, K.K. et al. (2001) J Biol Chem 276, 19276-85.
  6. Graves, J.D. et al. (1998) EMBO J 17, 2224-34.

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.