Product Specifications:
Item#1052: Recombinant tat HIV-1 Bal (E.coli)
Concentration: 1mg/ml
Mass/vial: 100ug
Volume/vial: See vial
Diluent: 50mM Tris, 0.1M NaCl, 2mM dtt, pH7.8
Purity: >95%
Stabilizer: None
Preservative: None
Storage: -75°C
Physical State: Frozen Liquid
Stability: At least 6 months at -75°C.
Application: ELISA, Western ELISA, Anti-tat Drug Screening, Immunization, Transcriptional Activation.
Description: Full length (101 amino acids) Recombinant HIV-1 Bal tat produced in the E.coli Expression System.
Purification: This protein was purified by solvent extraction, ion affinity and macro-molecular sieving to approx.95% purity, as determined by SDS-PAGE, reduced.
MW: approx.16kD.
Specificity: This protein binds to murine monoclonal antibodies of defined epitope specificity and rabbit and human serum polyclonal antibodies in ELISA and Western ELISA.
Biological Activity: Not Done
Endotoxin: Assay pending
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. Dilute tat stock solution in saline-phosphate buffer (150mM NaCl, 50mM sodiumphosphate, pH 6.5) immediately before use. Tat readily oxidizes in buffer solutions which may change its LTR- dependent transcriptional activation activity. Transcriptional activation assays with tat are performed in 1-10µg/ml range. ELISA and Western ELISA require tat in 10-100ng protein range.
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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已经试过的方法有：电吹风烘箱
将试的方法是：超声
大家有什么好建议，谢谢了！
祝大家元宵节快乐!
我用乙醇来提取，但是回收瓶中只有乙醇，那里出问题了呢？？？？
我们有个中药项目，需要采用乙酸乙酯萃取，有效成分分布在乙酸乙酯有机层中，现在我们采用旋转蒸发仪回收乙酸乙酯直到蒸发瓶中有效成分成蜂窝固体状才取出，但现在有一个问题，我如何判断蒸发瓶中蜂窝状固体可以刮出？目前我们实验室做的时候是根据蒸发瓶释放真空时蜂窝状固体不收缩的标准作为蒸干取出固体物的，但这个标准太粗放了，我也知道光靠旋转蒸发仪是不可能除尽有机溶剂，我们后续还有一个工艺可以除去蒸干固体残留的有机溶剂。我现在想知道有没有一些判断标准用于旋转蒸发仪固体蒸干可以取出来的标准可供参考啊？盼指教，谢谢
1.我利用氯仿萃取生物碱，需要将氯仿蒸干，我做的样品有非常多份，但是每份样品很少，用的氯仿也很少，每份只用十几毫升左右，但是旋转蒸发仪的蒸发瓶好大，而且一次只能蒸发一个样，太慢了，就算蒸发完了，取出生物碱的时候也不好取，由于我做的是定量试验，所以如果取出生物碱时有残留会影响实验结果，到底怎么办啊？有没有特制的小的蒸发瓶？或者特制的可以一次接好几个蒸发容器的接口？
2.请问恒温水浴锅可不可以用来做蒸发？由于是蒸发氯仿，担心没有回收装置会导致泄露中毒的问题，但是我的量比较少，又想冒险试试，真是焦头烂额了现在，大家把那个我出出主意吧！！！！
