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

PubChem ID

667639

Name

Piceatannol

Measurement

3.9μM - IC50

PubChem ID

122130844

Name

Staurosporine

Measurement

0.019μM - IC50

Clinical Relevance

Therapeutic Area:

Inflammation/Allergy

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

RELATED SOLUTIONS

Item: PP90