HIV-1 Protease, Functional (5x10μg)

Cat.No: FNK-05-013-5

Size: 5 x 10 µg

Storage: -80°C Avoid freeze-thaw cycles

Concentration: 0.2 mg/ml

Buffer: 10% glycerol, 20mM Tris, 20mM MES, 0.2M NaCl, 1mM EDTA, 1mM DTT, pH6.5

Purity: Over 90% by SDS-Page (CBB staining)

Reaction Buffer: 20 mM Tris-HCl (pH 6.8), 1 mM EDTA, 1 mM DTT, 0.1% Triton X-100, 10% Glycerol
 

Product Description

Full-size funcional recombinant HIV-1 protease purified from E.coli.

Application

1. Functional studies of HIV-1 Nef protein

2. SDS-page and Western blotting

3. ELISA

Background

HIV-1 protease is the aspartyl protease that mediates proteolytic cleavages of Gag and Gag-Pol polyproteins during or shortly after the release of the virion from the plasma membrane. Cleavages take place as an ordered, step-wise cascade to yield mature proteins. This process is called maturation. Displays maximal activity during the budding process just prior to particle release from the cell. Also cleaves Nef and Vif, probably concomitantly with viral structural proteins on maturation of virus particles. Hydrolyzes host EIF4GI and PABP1 in order to shut off the capped cellular mRNA translation. The resulting inhibition of cellular protein synthesis serves to ensure maximal viral gene expression and to evade host immune response.
 

Data Link

UniProt: P03367 (gag-pol), UniProt Q9YQ30 (HIV-1 Protease)
 

References

The HIV-1 strain and the recombinant protease has been described in the following references.

1. Adachi A et al “Production of acquired immunodeficiency syndromeassociated retrovirus in human and nonhuman cells transfected with an infectious molecular clone” J Virol 59: 284 -291(1986) PMID: 3016298

2. Saitoh A et al “Overproduction of human immunodeficiency virus type I reverse transcriptase in Escherichia coli and purification of the enzyme” Microbiol Immunol 34:509-521 (1990) PMID: 1699113

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. NOT FOR MILITARY USE.
 

Data Image

05-013, 05-012 HIV-1 Protease

Fig.1 SDS-PAGE analysis of purified HIV-1 protease.



Fig.2 Proteolytic processing of HIV-1 Gag p55 proprotein by HIV-1 protease in vitro 



As the substrate, recombinant Gag p55 (1 µg, BioAcademia 05-009 ) was used in 20 µl reaction volume. The reaction was carried by incubating at 37℃ for 3 h and stopped by adding SDS-PAGE sample buffer. 1; no protease, 2: 0.16 pg. 3; 1.6 pg. 4; 16 pg 5; 0.16 µg . 6; 1.6 µg protease. Note that two degradation bands are observed in the preparation of p55 substrate. In lane 4, p25 band is visible and in lane 5, p13 band is visible.

Fig.3 Purified HIV-1 protease as control antigen for Western blotting analysis with anti-HIV-1 protease antibody.


Fig.4  Dot blotting of HIV-1 protease by using anti-HIV-1 protease antibody



Fig.5 ELISA of HIV-1 protease with anti-HIV-1 protease antibody (BioAcademia 65-018). 



The antibody was used at dilutions indicated above. Purified Protease was spotted on wells. 
 

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FNK-05-013	HIV-1 Protease, Functional (10 µg)	GO!