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:
ebiomall.com
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如果PCR产物不是很纯，或者PCR扩增条带比较小，PCR产物前面又有较多引物二聚体时，用胶回收，其余用PCR产物纯化试剂盒。
pcr纯化试剂盒和胶回收试剂盒的区别：
PCR纯化试剂盒：是直接水溶解的PAC产物就可以回收，回收效率高，但是只适合单一条带需要纯化测序的时候使用。
PCR凝胶试剂盒：是在PCR产物是混合物，有多条杂带的情况下，先跑胶将杂带分离，然后在将所要的条带位置的胶切下回收，后者的回收效率低，但是很纯净。
胶回收试剂盒操作步骤：
配制琼脂糖EB凝胶,电泳以分离DNA片段。任何类型或等级的琼脂糖都可以使用。
电泳足够时间后，在紫外灯下小心地把所需的DNA的片段切下来。并尽量去除多余的凝胶。
称取空离心管的重量，切下带目的片段的凝胶装在1.5ml离心管中并称其重量，求出凝胶块的重量，近似地确定其体积。一般情况下，凝胶的密度为1g/ml，于是凝胶的体积与重量的关系可按下面换算：凝胶薄片的重量为0.2g 则其体积为0.2ml；加入等倍凝胶体积的Binding Buffer，把混合物置于55℃~65℃水浴中温浴7min至凝胶完全融化，其间每隔2-3分钟混匀一次；
转移700μl的DNA-琼脂糖溶液到一个HiBindTM DNA柱子，并把柱子装在一个干净的2ml收集管内，室温下，10,000×g离心1min，弃去液体。
将柱子重新套回收集管中，加300μl Binding Buffer至HiBind DNA 柱子中；室温下，10,000×g离心 1分钟，去弃滤出液；这一步相当关键，不要忽略此步。
将柱子重新套回收集管中，加入700μl SPW Wash buffer至HiBind DNA柱子中，室温下，10,000×g离心1分钟，去弃滤出液；注：SPW Wash buffer在使用前必须按瓶子标鉴要求用无水乙醇进行稀释。
将柱子重新套回收集管中，重复加入700μl SPW Wash buffer至HiBind DNA柱子中，室温下，10,000×g离心1分钟，去弃滤出液；
弃去液体，将空柱子重新套回收集管中，10,000×g离心1min以甩干柱基质残余的液体。
这步可以去除柱子基质上残余的乙醇，不要省略此步―――对得到好的DNA产量是十分重要的。
把柱子装在一个干净的1.5ml离心管上，加入30~50μl洗脱液或灭菌水上柱子膜上，10,000×g离心1分钟，离心管中的溶液就是纯化的DNA产物，保存于-20度。
它一般在医院、制药企业使用。
试剂盒使用示例： 试剂盒的产生正是为了使实验人员能够摆脱繁重的试剂配制及优化过程，所以试剂盒中一般配备有相应的使用说明书
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
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
将来可能会出现基于免疫方法的试剂盒，不过因为H7N9是个新病毒，爆发至今不足一月，这么快时间还来不及生产抗体，所以目前的检测试剂盒只能是PCR检测试剂盒。
查到了：
4月7日，上海之江生物科技有限公司官方微博称，该公司成功研制禽流感H7N9(2013)核酸测定试剂盒(荧光PCR法)，是针对国内此次H7N9病毒最早研制成功的产品，也是国内目前唯一供应的成品化试剂盒。
