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
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TEKLWVTVYY GVPVWKEATT TLFCASDAKA YDTEVHNVWA THACVPTDPN PQEVVLVNVT ENFNMWKNDM VEQMHEDIIS LWDQSLKPCV KLTPLCVSLK
110 120 130 140 150 160 170 180 190 200
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CTDLKNDTNT NSSSGRMIME KGEIKNCSFN ISTSIRGKVQ KEYAFFYKLD IIPIDNDTTS YKLTSCNTSV ITQACPKVSF EPIPIHYCAP AGFAILKCNN
210 220 230 240 250 260 270 280 290 300
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KTFNGTGPCT NVSTVQCTHG IRPVVSTQLL LNGSLAEEEV VIRSVNFTDN AKTIIVQLNT SVEINCTRPN NNTRKRIRIQ RGPGRAFVTI GKIGNMRQAH
310 320 330 340 350 360 370 380 390 400
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CNISRAKWNN TLKQIASKLR EQFGNNKTII FKQSSGGDPE IVTHSFNCGG EFFYCNSTQL FNSTWFNSTW STEGSNNTEG SDTITLPCRI KQIINMWQKV
410 420 430 440 450 460 470 480
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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.
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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).
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欧美国家是否有相关权威的抗生素使用规范？如果有，请提供，最好是中英对照的，相关网站也可以
伤,有必要使用含有DMSO冷冻保护剂。 DMSO能够快速穿透细胞膜进入细胞中,降低冰点、延缓冻存过程, 同时提高细胞内离子浓度,
减少细胞内冰晶的形成,从而减少细胞损伤。深低温时二甲基亚砜的细胞毒性受到抑制。复苏时动作要快，尽快洗掉二甲基亚砜，否则会造成对细胞严重的毒性。二
甲基亚砜（DMSO）是目前最好的细胞冻存保护剂，但也是一种以细胞毒性很大的化学试验剂。研究结果表明，培养液中DMSO浓度为10%时，细胞生长抑制
率近100%；1‰浓度时抑制率为35%，即使是0.04‰的浓度，DMSO对细胞的生长也有不利的影响。
1、血清：大部分人提及的Hyclone和Gibco，现在很多人对其血清的来源及目前市面上流行的真假好坏有怀疑，有人买到说好，有人说像是假的；也有人用国产的胎牛血清培养的，多提及兰州民海和四季青的，并且对兰州民海似乎支持者较多。对血清的选择还想说，不同厂家不同批次的血清都不同保证成分完全一致，对于使用者而言，可以从外观、培养方面了解血清的质量。
2、内皮生长因子：养内皮细胞都需加入生长因子，ECGS和ECGF用的较多，含量ECGS0.03-0.05mg/ml或ECGF50μg/ml，品牌提的最多的是Sigma的，但是Sigma的真心贵啊，另有些人提的最多的是罗氏，较便宜，75mg才2200RMB左右，性价比应该是比较高的，本人准备入手。
3、肝素：肝素在内皮细胞培养中的作用是抑制其他杂细胞的生长，尤其是平滑肌细胞的生长，使内皮细胞成为生长优势群，浓度不定，有几种说法：查阅书，15μg/ml；美国ATCC说明书，0.1mg/ml；另有50U/ml，肝素买的话大部分人也都推荐Sigma。
细胞冻存是细胞保存的主要方法之一。利用冻存技术将细胞置于-196℃液氮中低温保存，可以使细胞暂时脱离生长状态而将其细胞特性保存起来，这样在需要的时候再复苏细胞用于实验。而且适度地保存一定量的细胞，可以防止因正在培养的细胞被污染或其他意外事件而使细胞丢种，起到了细胞保种的作用。除此之外，还可以利用细胞冻存的形式来购买、寄赠、交换和运送某些细胞。 细胞冻存时向培养基中加入保护剂--终浓度5%．15%的甘油或二甲基亚砜(DMSO)，可使溶液冰点降低，加之在缓慢冻结条件下，细胞内水分透出，减少了冰晶形成，从而避免细胞损伤。 采用"慢冻快融"的方法能较好地保证细胞存活。标准冷冻速度开始为-1 到 -2℃/min，当温度低于-25℃时可加速，到-80℃之后可直接投入液氮内(-196℃)。
成纤维细胞生长抑制试剂盒—FibrOut™ 是由多种生化复合物组成的混合物、抗体或特殊的试剂。它能够抑制成纤维细胞过度增长或污染从而有效地增加靶细胞的产量。每种成纤维细胞生长抑制试剂盒都可定制到特定组织和特定细胞，经验证可与对应的PrimaCellTM系统高效协同工作，可为您带来每一类原代细胞培养的最佳结果。每种成纤维细胞生长抑制试剂盒都配备了最合适的缓冲液。
FibrOut™ 是一种包含几种生化复合物和试剂的完整系统，它在原代细胞培养中能抑制成纤维细胞增殖，从而促进靶细胞生长。每种成纤维细胞生长抑制试剂盒都可定制到特定组织和特定细胞，经验证可与对应原代细胞培养试剂盒-PrimaCellTM高效协同工作，可为您带来每一类原代细胞培养的最好结果。每种成纤维细胞生长抑制试剂盒都配备了最合适的缓冲液。成分包括 (1)胰蛋白酶；(2)胶原酶；(3)D-缬氨酸；(4)顺式羟基脯氨酸；(5)硫柳汞；(6)苯巴比妥；(7)系列血清替代品；(8)抗中胚层带抗体。
参考文献：
1. Cancer Res. 2010 May 1;70(9):3537-46，Epub 2010 Apr 20. Requirement of the NF-kappaB subunit p65/RelA for K-Ras-induced lung tumorigenesis. Bassères DS, Ebbs A, Levantini E, Baldwin AS.
2. Nat Commun. 2013;4:1795. Recruitment of mesenchymal stem cells into prostate tumours promotes metastasis. Jung Y, Kim JK, Shiozawa Y, Wang J, Mishra A, Joseph J, Berry JE, McGee S, Lee E, Sun H, Wang J, Jin T, Zhang H, Dai J, Krebsbach PH, Keller ET, Pienta KJ, Taichman RS.
3. Mol Biol Cell. 2012 Jul;23(14):2755-69. Calcium-calmodulin kinase I cooperatively regulates nucleocytoplasmic shuttling of CCTα by accessing a nuclear export signal. Agassandian M, Chen BB, Pulijala R, Kaercher L, Glasser JR, Mallampalli RK.展开
介绍了日本常用的药用辅料的安全性数据，类似美国的IIG，有较大参考价值，
举例
和名 ポリオキシエチレン（105）ポリオキシプロピレン（5）グリコール
英文名 Polyoxyethylene(105)Polyoxypropylene(5)Glycol
CAS9003-11-6
別名 PEP-101(110672)
収載公定書 薬添規(2003)
用途 界面活性剤，可塑剤，可溶（化）剤，コーティング剤，賦形剤，分散剤，崩壊剤，糖衣剤
■最大使用量
経口投与 400mg
以下については該当文献なし.
■単回投与毒性
■反復投与毒性
■遺伝毒性
■癌原性
■生殖発生毒性
■局所刺激性
■その他の毒性
■ヒトにおける知見
■引用文献
下面是连接，版头有搜索入口，支持日文检索
http://www.jpec.gr.jp/detail=normal&date=safetydata/data.html
