Trypsin (Non-Selective) Human Chymotrypsin Serine Peptidase Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Trypsin is a serine proteinase and known as an essential food-digestive enzyme. Four trypsin (or trypsinogen) genes have been characterized in humans. Trypsins 1, 2, and 3 have been detected in human pancreatic juice. Trypsin 2 is expressed by vascular endothelial cells around gastric tumors and in patients with disseminated intravascular coagulation. The trypsin 4 gene is expressed in the human brain. Trypsins activate protease activated receptors (PARs) 1-4, proelastase, pro-MMP-2, pro-MMP9 and others. These facts suggest broad distribution and function of trypsin in normal tissues. Trypsin has been detected in pancreatic cancers and is associated with hereditary pancreatitis.
Family:
Protease
Sub Family:
Chymotrypsin
Class:
Serine Peptidase
Accession Number:
NM_002769
Organism:
Human
Construct Details:
Endogenous
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Human pancrease trypsin is used. Test compound and/or vehicle is preincubated with 0.016 nM enzyme‡ in modified HEPES buffer pH 7.5 for 15 minutes at 25⁰C. The reaction is initiated by addition of 20 μM Z-Gly-Pro-Arg-AMC for another 60 minute incubation period. Determination of the amount of AMC formed is read spectrofluorimetrically at 360 nm/465 nm. Compounds are screened at 10 μM.
Substrate:
20 µM Z-Gly-Pro-Arg-AMC
Incubation:
60 min at 25°C
Control Inhibitor:
Camostat mesylate
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
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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