Pepsin Human Aspartic Peptidase Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Pepsin produced in the mucosal lining of the stomach acts to degrade protein. Pepsin is one of three principal protein-degrading, or proteolytic, enzymes in the digestive system, the other two being chymotrypsin and trypsin. It is activated by stomach acid. Inhibition of pepsin may be useful to prevent inflammation after gastrointestinal surgery.
Family:
Protease
Sub Family:
Pepsin
Class:
Aspartic Peptidase
Protein Name:
Pepsinogen
Uniprot Number:
P0DJD7
Protein Aliases:
Pepsinogen-4
Gene Name:
PGA4
Gene ID:
643847
Gene Aliases:
pepsin A
Accession Number:
NM_001079808
Organism:
Human
Construct Details:
full length recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Platform:
Plate reader
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Human recombinant pepsinogen A expressed in mouse myeloma cells is used. Test compound and/or vehicle is preincubated with 10.9 ng/ml enzyme in sodium citrate buffer pH 4.0 for 15 minutes at 25°C. The reaction is initiated by addition of 3 mM MOCAc-Ala-Pro-Ala-Lys-Phe-Phe-Arg-Leu-Lys(Dnp)-NH2 for another 30-minute incubation period. Determination of the amount of MOCAc-Ala-Pro-Ala-Lys-Phe formed is read spectrofluorimetrically at 325 nm/400 nm. Compounds are screened at 10 µM.
Substrate:
3 µM MOCAc-Ala-Pro-Ala-Lys-Phe-Phe-Arg-Leu-Lys(Dnp)-NH2
Incubation:
30 min at 25°C
Control Inhibitor:
Pepstatin
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).
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Gastroenterology
Adverse / Beneficial Action:
Inhibition of pepsin may impair protein digestion and assimilation as well as prevent inflammation after gastrointestinal surgery.
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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