HIV-2 Protease Activity assay kit utilizes the ability of active HIV- 2 Protease to cleave a synthetic peptide substrate to release the free fluorophore which can be easily quantified (Ex/Em = 330/450 nm) using a fluorometer or fluorescence microplate reader. This assay is simple, rapid and can detect HIV-2 protease activity in samples and of purified enzyme.
Human Immunodeficiency Virus (HIV) is the cause of the Acquired Immunodeficiency Syndrome (AIDS). Infection with HIV-2 virus causes immunosuppression, as well as AIDS, characterized by the same signs, symptoms and opportunistic infections as HIV-1. However, HIV-2- associated AIDS may often be associated with lower viral load levels than HIV-1. Aspartyl HIV-2 Protease (retropepsin) is an essential enzyme for the life-cycle of HIV virus. Expressed as a part of the Gag-Pol polyprotein complex in the infected cells, it is auto-catalytically released upon formation of an immature viral particle. The enzyme subsequently cleaves the other parts of viral polyproteins resulting in the maturation of the virus. In HIV-infected patients the enzyme is subjected to intensive mutagenesis and mutants resistant to applied medicines are produced as a result of the selection pressure.
Detect activity of recombinant HIV-2 proteaseDetermine activity of purified HIV-2 protease
Store kit at -80°C, protected from light. Briefly centrifuge small vials at low speed prior to opening. Read the entire protocol before performing the assay.HIV-2 Protease Assay and Dilution Buffers: Bring to room temperature before use. Store at -20°C.HIV-2 Protease: Add 20 µl of HIV Protease Dilution Buffer to the vial. Aliquot and store at -80°C. Avoid repeated freeze/thaw.
HIV-2 Protease Assay Buffer: 25 mlHIV-2 Protease Dilution Buffer: 1 mlHIV-2 Protease Substrate: 0.2 mlHIV-2 Protease (Positive Control): 20 μlFl. Standard (10 mM in DMSO): 20 μ
Fluorescence (Ex/Em = 330/450 nm)
Compatible Sample Types:
Features & Benefits:
• Simple, sensitive assay• Fluorometric detection