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7752
HTScan® Jak2 Kinase Assay Kit

HTScan® Jak2 Kinase Assay Kit #7752

This product is discontinued

Kinase Assay - Radiometric Image 1

Figure 1. Jak2 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: Tetra (LRRWSLG), 5 µg/50 µl, recombinant Jak2: 50 ng/50 µl.

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Kinase Assay - DELFIA Image 2

Figure 3. Dose dependence curve of Jak2 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1305) by Jak2 kinase. In a 50 µl reaction, increasing amounts of Jak2 and 1.5 µM substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Kinase Assay - DELFIA Image 3

Figure 5. Staurosporine inhibition of Jak2 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of Jak2 substrate peptide (#1305) by Jak2 kinase. In a 50 µl reaction, 100 ng Jak2, 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.)

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Kinase Assay - DELFIA Image 4

Figure 4. Peptide concentration dependence of Jak2 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) to detect phosphorylation of substrate peptide (#1305) by Jak2 kinase. In a 50 µl reaction, 100 ng of Jak2 and increasing concentrations of substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Kinase Assay - DELFIA Image 5

Figure 2. Time course of Jak2 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) to detect phosphorylation of Jak2 substrate peptide (#1305) by Jak2 kinase. In a 50 µl reaction, 100 ng Jak2 and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Product Includes Quantity (with Count) Solution Color
DTT (1000X) 1 x 80 µl
ATP (10 mM) 9804 1 x 1 ml

The kit provides a means of performing kinase activity assays with recombinant murine Jak2 kinase. It includes active Jak2 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:

FLT3 (Tyr589): 1,945 Daltons, GST-Jak2 Kinase: 71 kDa Daltons.

Peptide Core Sequence:

NEY*FY*V

Members of the Janus family of tyrosine kinases (Jak1, Jak2, Jak3, and Tyk2) are activated by ligands binding to a number of associated cytokine receptors (1). Upon cytokine receptor activation, Jak proteins become autophosphorylated and phosphorylate their associated receptors to provide multiple binding sites for signaling proteins. These associated signaling proteins, such as Stats (2), Shc (3), insulin receptor substrates (4), and focal adhesion kinase (FAK) (5), typically contain SH2 or other phospho-tyrosine-binding domains.

  1. Leonard, W.J. and O'Shea, J.J. (1998) Annu. Rev. Immunol. 16, 293-322.
  2. Darnell, J.E. (1997) Science 277, 1630-5.
  3. VanderKuur, J. et al. (1995) J Biol Chem 270, 7587-93.
  4. Argetsinger, L.S. et al. (1995) J Biol Chem 270, 14685-92.
  5. Zhu, T. et al. (1998) J Biol Chem 273, 10682-9.
Entrez-Gene Id
3717
Swiss-Prot Acc.
O60674
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.

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