TACE Human Astacin/Adamalysin Metallo Peptidase Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Tumor necrosis factor-alpha (TNF-a) converting enzyme (TACE/ADAM-17) is responsible for the release of TNF-a, a potent proinflammatory cytokine associated with many chronic debilitating diseases such as rheumatoid arthritis and multiple sclerosis. TNF-a also induces angiogenesis, promotes fibroblast proliferation and can combine with receptors on selected tumor cells to induce cell lysis. TACE is also implicated in amyloid precursor protein secretion.
Family:
Protease
Sub Family:
Astacin/Adamalysin
Class:
Metallo Peptidase
Protein Name:
TACE
Uniprot Number:
P78536
Gene Name:
ADAM17
Gene ID:
6868
Gene Aliases:
cSVP|CD156B
Accession Number:
NM_003183
Organism:
Human
Construct Details:
Fragment (215-671), Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Human recombinant TACE expressed in Sf21 insect cells is used. Test compound and/or vehicle is preincubated with 25 ng/ml‡ enzyme in modified Tris-HCl buffer pH 9.0 for 15 minutes at 25°C. The reaction is initiated by addition of 10 μM Mca-Pro-Leu-Ala-Gln-Ala-Val-Dpa-Arg-Ser-Ser-Ser-Arg for another 30 minute incubation period. Determination of the amount of Mca-Pro-Leu-Ala-Gln-Ala formed is read spectrofluorimetrically at 340 nm/400 nm. Compounds are screened at 10 μM.
Substrate:
10 µM Mca-P-L-A-Q-A-V-Dpa-R-S-S-S-R-NH2
Incubation:
30 min at 25°C
Control Inhibitor:
Ilomastat
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:
Oncology/Immuno-Oncology
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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