Biological Information
Background Information:
ZA70 (Tyrosine Protein kinase ZAP-70, 70 kDa zeta-associated protein, Syk-related tyrosine kinase) is a cytosolic protein tyrosine kinase (PTK) and a member of the Syk family of proteins. ZA70 is expressed in human and murine thymocyte subsets, B cells, and peripheral T cell subsets and associates with zeta-chain to undergo tyrosine phosphorylation following TCR stimulation. ZA70 inhibitors may be useful as immunosuppressants.
Family:
Kinase
Sub Family:
TK
Class:
Protein Tyrosine
Protein Name:
ZAP70
Uniprot Number:
P43403
Protein Aliases:
tyrosine-protein kinase ZAP-70
Gene Name:
ZAP70
Gene ID:
7535
Gene Aliases:
ZAP-70|STD
Accession Number:
NM_001079
Organism:
Human
Construct Details:
Full Length, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometry
Measured Response:
Scintillation
Testing Information
Procedure Summary:
Human recombinant protein kinase ZAP-70 expressed in insect cells is used. Test compound and/or vehicle is preincubated with 0.45 U/ml enzyme‡ for 15 minutes at 37°C in modified HEPES buffer pH 7.4. The reaction is initiated by addition of 0.2 mg/ml poly(Glu:Tyr), 10 µM ATP containing 0.25 µCi [γ32P]ATP for 30 minutes and terminated by further addition of 3% H3PO4. An aliquot is counted to determine the radioactivity of [32P]Poly(Glu:Tyr). Compounds are screened at 10 µM.
Substrate:
200 µg/ml Poly(Glu:Tyr)
ATP Concentration:
10 uM
Incubation:
30 min at 37°C
Control Inhibitor:
Staurosporine
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Inflammation/Allergy
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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