HIV-1 Protease, functional
05-013 10 µg,05-013-550 µg

Storage: Store at -80℃. Avoid freeze-thaw cycles.
Product: Full-size functional recombinant HIV-1 protease purified from E. coli
Applications
1.

Functional studies such as screening for inhibitors of HIV-1 protease for AIDS drug
discovery

2.

Antigen for anti-HIV 1 antibody for Western blotting and ELISA

3.

Control as HIV 1 protease for SDS=PAGE
Other applications have not been tested.

Stock solution: 0.2 mg /ml in 20 mM Tris, 20 mM MES, 0.2 M NaCl, 1 mM

EDTA, 1mM DTT, 10% Glycerol, pH 6.5
Reaction buffer: 20 mM Tris-HCl (pH 6.8), 1 mM EDTA, 1 mM DTT, 0.1% Triton X-100,10% Glycerol
Background: HIV-1 protease is the aspartyl protease that mediates proteolytic cleavagesof Gag and Gag-Pol polyproteins during or shortly after the release of the virion from theplasma membrane. Cleavages take place as an ordered, step-wise cascade to yield
mature proteins. This process is called maturation. Displays maximal activity during thebudding 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 mRNAtranslation. 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 thefollowing references.
1. Adachi Aet al “Production of acquired immunodeficiency syndrome-associatedretrovirus in human and nonhuman cells transfected with an infectious molecularclone” 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: 16991

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kDa

1

23

kDa

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

75503725201510M

123456kDa

7550p553725p2420p1715p1310ProteaseFig.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. Notethat two degradation bands are observed in the preparation of p55 substrate. In lane 4, p25 band isvisible and in lane 5, p13 band is visible.
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Fig.3 Purified HIV-1 protease as control antigen for Western
blotting analysis with anti-HIV-1 protease antibody.
1: Purified HIV-1 protease (1 ng )2: Extract of MT4 cells
3.: Extract of MT4 cells infected with HIV-1 (LAI strain)
The anti-HIV-1 protease antibody (BioAcademia 65-018) wasused at 1/2,000 dilution.

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

Anti-HIV-1 protease antibody (BioAcademis 65-018) was used at 1/2,000 dilution. As second
antibody, goat anti-rabbit IgG antibody conjugated with HRP was used at 1/5,000 dilution.

Fig.4 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.
Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
BioAcademia,Inc.
0Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp

https://www.bioacademia.co.jp/en/