HIV-1 Protease Virus Aspartic Peptidase Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
When viral RNA is translated into a polypeptide sequence, that sequence is assembled in a long chain that includes several individual proteins (reverse transcriptase, protease, integrase). Before these enzymes become functional, they must be cut from the longer polypeptide chain. Viral protease cuts the long chain into its individual enzyme components, which then facilitate the production of new viruses. HIV-1 protease is a novel aspartic protease that functions to cleave the nascent polyproteins during viral replication. HIV protease inhibitors can lower the viral load carried by AIDS patents.
Family:
Protease
Class:
Aspartic Peptidase
Organism:
Virus
Construct Details:
Full Length, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Recombinant HIV protease expressed in E. coli is used. Test compound and/or vehicle is preincubated with 4 μg/ml of enzyme‡ in modified disodium acetate buffer pH 4.7 for 15 minutes at 25°C. The reaction is initiated by addition of 30 μM Anthranilyl-HIV protease substrate for another 5 minute incubation period. Determination of the amount of Anthranilyl-tripeptides formed is read spectrofluorimetrically at 400 nm. Compounds are screened at 10 μM.
Substrate:
30 µM Anthranilyl-HIV protease substrate
Incubation:
5 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).
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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