JAK2-JH2 Pseudokinase Domain (W659A, W777A, F794H) Inhibitor Screening Assay Kit

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Background: Janus kinases (JAKs) are a family of intracellular nonreceptor tyrosine kinases including JAK1, JAK2, JAK3 and Tyk2, that has been recognized as an important modulator in inflammatory processes. JAKs contain a catalytically inactive pseudokinase regulatory domain (JH2) as well as an active kinase domain (JH1). Selective inhibition of one specific JAK is a challenging task since the enzymes share high homology in the active site of JH1. Recent reports demonstrate that the pseudokinase domain (JH2) could provide an ideal allosteric site for selective inhibitors.

Contraindications: Keep final DMSO concentration at or below 1%.

Description: The JAK2 JH2 Pseudokinase Domain Inhibitor Screening Assay Kit is designed for screening and profiling small molecules that displace the fluorescently labeled probe (JH2 probe 1) from the JH2 domain of JAK2. The assay is a fluorescence polarization (FP) assay based on the competition of the test compound with the JH2 probe for binding to purified JAK2 JH2.

JH2 probe 1 is a small molecule probe that binds to the ATP-binding site of JAK2 pseudokinase domain JH2. The probe has been labeled with a fluorescent tracer, so that binding of the probe to much larger JH2 results in noticeable increases in fluorescence polarization compared to the unbound fluorescent probe (depolarized). Competition between a test drug and the probe results in changes in polarization. Using this kit, only one simple step on a microplate is required for screening. The JH2 probe 1 is incubated with a sample containing JAK2 JH2 to produce a change in fluorescent polarization that can then be measured using a fluorescence microplate reader capable of measuring fluorescence polarization.



Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Supplied As: This kit comes in a convenient 384-well format, with enough recombinant human JAK2 JH2 and fluorescently labeled ATP binding site tracer (JH2 probe 1) for 384 reactions.

Warnings: Avoid freeze/thaw cycles.

Biosafety Level: Not applicable (BSL-1)

References: 1. Wrobleski, S. T., et al. J. Med. Chem. 2019, 62(20): 8973-8995.
2. Hubbard, S. R. "Mechanistic insights into regulation of JAK2 tyrosine kinase." Frontiers in endocrinology 2018, 8: 361.
3. Newton, Ana S., et al. "JAK2 JH2 fluorescence polarization assay and crystal structures for complexes with three small molecules." ACS medicinal chemistry letters 2017, 8(6): 614-617.
4. Yamaoka, Kunihiro, et al. "The Janus Kinases (Jaks)." Genome biology 2004, 5(12): 1-6.

• Artikelnummer: BPS78208
• Hersteller: BPS Bioscience
• Hersteller Artikelnummer: 78208
• Green Labware: Nein
• Verpackungseinheit: 384 rxns.
• Mengeneinheit: PAK