Product Specifications:
Item# 1001: Recombinant HIV-1 IIIB Envelope Glycoprotein gp120 (Baculovirus)
Concentration: See Vial
Mass/vial: 100ug
Diluent: PBS
Purity: >95%
Stabilizer: None
Preservative: None
Storage: -75°C
Physical State:Frozen Liquid
Stability: At least 2 years at -75°C.
Applications: In-Vitro Diagnostics, T-Cell Activation, CD4 Binding, Immunization, Antibody screen
Description: Full length Recombinant HIV-1 IIIB gp120 glycoprotein produced in the Baculovirus Expression System. Authentic N-C termini, no tags
Purification: This protein is purified by immune-affinity chromatography to >95% purity as determined by SDS-PAGE, reduced.
Specificity: This protein binds to murine monoclonal antibodies of defined epitope specificity and binding to HIV-1 converted human serum polyclonal antibodies in ELISA and Western ELISA.
Biological Activity: This protein binds to human T-cell receptor CD4 in ELISA and Western ELISA as determined by CD4/gp120/Anti-gp120 mAb-peroxidase capture ELISA. This protein activates human T-Lymphocytes (CD4+, CD4-), in vitro, as measured by RNA synthesis during G0 to G1 transition phase of antigen-binding competent Cells.
Application and Instruction for use
Recommended concentrations for use are approximate values. A dose dependent response assay should be performed to determine the optimal concentration for use in specific applications.
ELISA and Western ELISA require 10-100ng protein depending on the nature and affinity of the detection reagent. HIV-converted human serum polyclonal antibodies yield titers of 1:1000 or greater at 1-10ng of immobilized protein under standard ELISA conditions.
What is gp120?
gp120 - HIV-1 virus envelope protein- is derived from gp120 and gp41components of envelope gp160. gp120 (481 amino acids) name comes from it"s molecular weight - 120 kDa. gp120 is essential for entry into cells as it plays a vital role in the infection process. Full length gp120 is a highly glycosylated and modified protein composed of roughly 55% amino acids and 50% carbohydrates by weight.
Gp120 interaction:
Gp120 interacts with the CD4 receptor and chemokine co-receptors (CCR5 and CXR4). gp120 binding to CD4 induces conformational changes in gp120 and gp41 that leads to the fusion of the viral membrane with the host cell membrane.
Protein Sequence:
HIV-1 (HXB2):
10 20 30 40 50 60 70 80 90 100
| | | | | | | | | |
TEKLWVTVYY GVPVWKEATT TLFCASDAKA YDTEVHNVWA THACVPTDPN PQEVVLVNVT ENFNMWKNDM VEQMHEDIIS LWDQSLKPCV KLTPLCVSLK
110 120 130 140 150 160 170 180 190 200
| | | | | | | | | |
CTDLKNDTNT NSSSGRMIME KGEIKNCSFN ISTSIRGKVQ KEYAFFYKLD IIPIDNDTTS YKLTSCNTSV ITQACPKVSF EPIPIHYCAP AGFAILKCNN
210 220 230 240 250 260 270 280 290 300
| | | | | | | | | |
KTFNGTGPCT NVSTVQCTHG IRPVVSTQLL LNGSLAEEEV VIRSVNFTDN AKTIIVQLNT SVEINCTRPN NNTRKRIRIQ RGPGRAFVTI GKIGNMRQAH
310 320 330 340 350 360 370 380 390 400
| | | | | | | | | |
CNISRAKWNN TLKQIASKLR EQFGNNKTII FKQSSGGDPE IVTHSFNCGG EFFYCNSTQL FNSTWFNSTW STEGSNNTEG SDTITLPCRI KQIINMWQKV
410 420 430 440 450 460 470 480
| | | | | | | |
GKAMYAPPIS GQIRCSSNIT GLLLTRDGGN SNNESEIFRP GGGDMRDNWR SELYKYKVVK IEPLGVAPTK AKRRVVQREK R
Our product gp120 and other envelope proteins (gp1203B, gp120MN, gp120YU2, gp120 ADA, gp120C) are used by research insitutes, pharmaceuticals, universities, and reagents repositories world wide. These envelope proteins are classified as M-tropic, T-tropic, and M/T-tropic viruses infecting M,T, M/T cells respectively.
These products are available in bulk to measure regents repositories all over the world.
Safety Study of rgp120/HIV-1IIIB Vaccine
Recent evidence suggests that gp120 is the HIV-1 protein with the greatest potential as a vaccine against HIV-1 infection. The gp120 envelope protein may be produced by recombinant DNA technology, and studies have shown that the vaccine is capable of eliciting neutralizing antibody activity in both rodents and nonhuman primate species.
Official Study Title: A Phase I Study of the Safety and Immunogenicity of rgp120/HIV-1IIIB Vaccine in Healthy Adult Subjects (NOTE: Study Extended ONLY for Subjects Who Have Previously Received rgp120/HIV-1IIIB or rgp120/HIV-1MN on VEU 006 or VEU 006 Rollover Study)
Disease: HIV Infections
Treatment: Biological: rgp120/HIV-1IIIB and Biological: rgp120/HIV-1MN
ORIGINAL DESIGN: Twenty-eight subjects will be randomized to receive 100 or 300 mcg rgp120/HIV-1IIIB vaccine (gp120 vaccine) or matching placebo. For each dose level, 10 subjects will receive vaccine and four subjects will receive matching placebo. Injections are given intramuscularly at 0, 4, and 32 weeks. Each subject receiving treatment at the lower dose level must be monitored for unacceptable toxicity for at least 2 weeks following the initial immunization before his or her second dose is administered and before treatment at the higher dose level begins. Subjects are followed for at least 12 months.
AMENDED 11/17/93: Selected subjects from VEU 006 or VEU 006 Rollover study will receive two injections of MN rgp120 vaccine, administered 28 days apart beginning 10-16 months after their last injection. Eight additional clinic visits will be required. Subjects are followed for at least 6 months.
_________________________________________________________________________
Glossary
Gene and Gene Products
Structural Proteins: Structural proteins – the products of gag, pol and env genes, which are essential components of the retroviral particle.
Regulatory Proteins: Regulatory proteins – tat and rev proteins of HIV/SIV and tax and rex proteins of HTLVs; essential for viral expression in infected cells.
Accessory Proteins: Accessory proteins – additional (non-regulatory) virion – and non virion-associated proteins produced by HIV/SIV retroviruses: vif, vpr, vpu, vpx, and nef. Although, the accessory proteins are not necessary for viral propagation in tissue culture, they have been conserved in the different isolates; this conservation and experimental observations suggest that their role in vivo is very important.
gag
gag – group-sepecifc antigens or capsid proteins; the precursor is the p55 myristoylated protein, which is processed to p17 (Matrix) p24 (Capsid) and p7 (NucleoCapsid) proteins by the viral protease. Other small proteins are generated from the gag polyprotein.
pol
pol – (p66) generates the viral enzymes protease (p11), reverse transcriptase (p51), endonuclease and integrase (p32) after the processing of a gag-pol precursor polyprotein by the viral protease; gag-pol precursor is produced by ribosome frameshifting.
env
env – viral glycoproteins produced as a precursor (gp160) and processed to the external glycoprotein (gp120) and the transmembrane glycoprotein (gp41). The mature proteins are held together by noncovalent interactions; as a result substantial amount of gp120 is released extracellularly. The external glycoprotein (gp120) contains the binding site for the CD4 receptor.
tat
tat – transactivator of HIV gene expression; one of the two necessary viral regulatory factors (tat and rev) for HIV gene expression. Two forms are known, tat-1 exon (minor form) of 72 amino acids, and tat-2 exon (major form) of 86 amino acids. The electrophoretic mobility of these two forms in SDS gels is anomalous; they are approximately 16 kD and 14 kD in weight. Low levels of both proteins are found in persistently infected cells. tat is localized primarily in the nucleolus/nucleus; it acts by binding to the TAR RNA element and activating transcription from the LTR promoter. Post-transcriptional effects of tat have been postulated.
rev
rev – the second necessary regulatory factor for HIV expression. A 19 kD phosphoprotein localized primarily in the nucleolus/nucleus, rev acts by binding to RRE and promoting the nuclear export, stabilization and utilization of the viral mRNAs containing RRE.
vif
vif – viral infectivity factor, typically 23 kD; required for the efficient transmission of cell-free virus in tissue culture. In the absence of vif, the produced viral particles are defective, while the cell-to-cell transmission of virus is not affected significantly. It has been reported that the cellular localization is in the Golgi (vif is not found in the virion).
nef
nef – approximately 27 kD non-virion protein found in the cytoplasm of infected cells. Potentially myristoylated and associated with the inner plasma membrane. One of the first HIV proteins to be produced in the infected cells, it is the most immunogenic of the accessory proteins and may be used in the future for diagnosis and staging of the disease. NEF is dispensable and probably suffers counter-selection during ex vivo viral propagation in vivo. Recent evidence suggests that SIV nef is required for viral propagation in vivo.
vpr
vpr – virion-associated protein of unknown function found in HIV-1, HIV-2, SIVmac, and SIVmnd; typically 15 kD. May be homologous to vpx. Also called “rap” for rapid.
vpu
vpu – protein that promotes extracellular release of viral particles. Found only in HIV-1. Integral membrane phosphoprotein of 16kd; similar to M2 protein of influenza virus. It may be involved in env maturation. It is not found in the virion.
vpx
vpx – virion protein of 12 kD found only in HIV-2 infection. (vpx may have some homology with vpr).
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
2乘SDS-PAGE上样缓冲液（20ml体系）
1.0mol/L Tris-HCL (pH 6.8) 2ml
1.0mol/L DTT 4ml
SDS 0.8g
溴芬蓝 0.04g
甘油 4ml
dd水 定容至20ml
4℃冰箱保存，可以不分装，我们实验室一般用三个月以上
先配1.0mol/L的DTT,双蒸无菌水溶解4℃冰箱保存,配上样缓冲液的时候加入就可以了,都混匀保存也没问题的
蛋白电泳上样缓冲液包括2%SDS , 0.1%溴酚蓝 10%甘油,SDS是保证样品中的所有蛋白带电荷一致，减少电荷对电泳结果的影响。还原剂是让二硫键处于断开状态，保证蛋白分子的线性.
溴酚蓝作用是指示上样蛋白的跑胶时标记的，甘油是增加上样重量的，以免漂浮，煮沸是使蛋白变性的永久保存
加样缓冲液的主要作用是使PCR产物与其混合,
使DNA沉于加样孔的底部,防止DNA跑出来.
