Revision 1

#7752Store at -80C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
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UniProt ID:

#O60674

Entrez-Gene Id:

3717

Product Information

Storage

Antibodies are supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Do not aliquot the antibodies. Peptides are supplied at 6 µM in 0.001% DMSO. Enzymes are supplied in 50 mM Tris-HCL (pH 8.0), 100 mM NaCl, 5 mM DTT, 15 mM reduced glutathione and 20% glycerol. Store at –80°C.
Keep enzymes on ice during use.
Avoid repeated freeze-thaw cycles.

Source / Purification

The GST-Jak2 Kinase fusion protein was produced using a baculovirus expression system with a construct expressing a fragment of murine Jak2 (Lys752-Val1129) (GenBank accession No. L16956) with an amino-terminal GST tag. The protein was purified by one-step affinity chromatography using glutathione-agarose.

Product Description

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.

Background

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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。
Revision 1
#7752

HTScan® Jak2 Kinase Assay Kit

HTScan® Jak2 Kinase Assay Kit: Image 1 Expand Image
图 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. 在 50 µl 反应体积中，每个反应孔在室温下用数量递增的 Jak2 和 1.5 µM 底物肽反应 30 分钟。（DELFIA® 是 PerkinElmer, Inc. 的注册商标）
HTScan® Jak2 Kinase Assay Kit: Image 2 Expand Image
图 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. 在 50 µl 反应体积中，每个反应孔在室温下用 100 ng Jak2、1.5 µM 底物肽、20 µM ATP 和数量递增的 staurosporine 反应 30 分钟。（DELFIA® 是 PerkinElmer, Inc. 的注册商标）
HTScan® Jak2 Kinase Assay Kit: Image 3 Expand Image
图 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. 在 50 µl 反应体积中，每个反应孔在室温下用 100 ng Jak2 和浓度递增的底物肽反应 30 分钟。（DELFIA® 是 PerkinElmer, Inc. 的注册商标）
HTScan® Jak2 Kinase Assay Kit: Image 4 Expand Image
图 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. 在 50 µl 反应体积中，每个反应孔用 100 ng Jak2 和 1.5 µM 底物肽。（DELFIA® 是 PerkinElmer, Inc. 的注册商标）
HTScan® Jak2 Kinase Assay Kit: Image 5 Expand Image
图 1. 使用以下反应条件在放射性同位素滤器结合测定中测量 Jak2 激酶活性：60 mM HEPES-NaOH、pH 7.5、3 mM MgCl2、3 mM MnCl2、3 µM 原钒酸钠、1.2 mM DTT、ATP（变量）、2.5 µg/50 µl PEG20.000，底物：Tetra (LRRWSLG)、5 µg/50 µl、重组 Jak2：50 ng/50 µl。
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Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。