Product Specifications:
Item# 1008: Recombinant HIV-1 nef (E.coli)
Concentration: See vial
Mass/vial: 100 ug
Volume/vial: 100ul
Diluent: 50mM Tris, pH8, 150mM NaCl
Purity: >95%
Stabilizer: None
Preservative: None
Storage: -75°C
Physical State: Frozen Liquid
Stability: Minimum 12 months at -75°C.
Application: Diagnostics, ELISA/ Western ELISA, Drug Screening, Immunological Studies.
Description: Full length Recombinant HIV-1 (LAV) nef produced as a GST fusion protein in E.coli.
Purification: This recombinant protein is purified by Glutathion -affinity chromatography and the purified protein is thrombin-cleaved to produce nef of >95% purity, as determined by SDS-PAGE, reduced.
Specificity: This protein shows reactivity with murine monoclonal antibodies and human polyclonal antibodies in ELISA and Western ELISA.
Biological Activity: Not done.
Application and Instructions 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 1-10ng protein depending on the nature and affinity of the test antibody.
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).
Related research paper:
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从1～2个T75培养皿的感染细胞培养液中纯化腺病毒
产品简介
本系列试剂盒可快速，高效地从腺病毒感染的细胞培养液中分离纯化腺病毒，相比于传统CsCl超速离心的腺病毒病毒纯化方式（需要24个小时才能完成纯化），本试剂盒可以在1个小时内完成病毒纯化，操作简单，快速，纯化率高，纯化到的病毒颗粒可直接用于下游实验，如细胞和动物感染。
产品特点
?操作简单，快速，可以在1个小时内完成病毒纯化，不依赖于超速离心操作。
?纯化效率高：小量纯化，可从1~2个T75瓶培养的细胞培养液中纯化病毒颗粒高达1x1012VPs
?每个纯化柱，可以重复利用一次，用于纯化相同种类的腺病毒。
保存条件
纯化柱和脱盐柱保存于4℃，其它组分室温保存。
试剂盒组分
Catalog#
V1160-01
Notes
Preps
10
MiniColumns
5
Canbeusedtwice
(Storeat4°C)
Press-OnCap
5
StoreatRT
DesaltingTube*
1
Canberegenarated
(Storeat4°C)
15mLCollectionTube
10
StoreatRT
10xWashBuffer
30mL
StoreatRT
2xElutionBuffer
30mL
StoreatRT
RegenerationBuffer
30mL
StoreatRT
联系方式：
Biomiga(中国）
电话：0573-82651206
QQ:441931287
联系人：张小姐
病毒液通过pre-filterdisc的时候是不是要制造一个真空环境产生压力差使液体通过？这个时候是用自备的针管还是试剂盒配的5毫升的针管啊？
试剂里面含有的是病毒的抗原，现在基本都是人工合成或提取的，跟病毒完全是两回事
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