Product Specifications:
Item#1009: Recombinant p31 HIV-1
Concentration: See vial
Mass/vial: 100ug
Volume/vial: See vial
Diluent: PBS, 0.05% Sarcosyl
Purity: >95%
Stabilizer: None
Preservative: None
Storage: -75°C
Physical State: Frozen Liquid
Stability: Minimum 12 months at -75°C
Application: ELISA, Western ELISA, Immunological Studies, Drug Screening.
Description: Full length HIV-1 endonuclese (p31) expressed in the the E.coli expression system.
Purification: This protein is purified by solvent extraction and preparative electrophoresis to >98% purity, as determined by SDS-PAGE, reduced.
Approx. Molecular Weight: 36kD.
Specificity: This protein binds to human anti-protease polyclonal antibodies and murine monoclonal antibodies as determined ELISA and Western ELISA
Biological Activity: Not determined.
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 10-100ng 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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2：冰满探头调整太靠近冰格，顺时针调整180度就好了，也有可能冰满探头坏了。需要更换。
3：水位浮球开关坏，更换。
不过制冰机里的冰不可以，那冰怎么得到呢？
protocol上面说在离心菌液的时候离心管里面要有1/2的冰啊，这个冰应该是碎冰而不是直接冻出来的整块的冰吧。
大家是怎么得到的呢？谢谢！！
概括了以下四点:
1.在用完制冰机后需要将冰箱内的冰块清理干净。清洗制冰机时应关掉电源，不要用水管直接对准机身冲洗，用清水，加入适量中性洗涤剂(严禁用酸性、碱性等腐蚀性溶剂)擦拭机器围板和内胆，再用软布擦干净。
2.制冰机必须两个月旋开进水软管管头，清洗进水阀滤网，避免水中砂泥杂质堵塞进水口，而引起进水量变小，导致不制冰。
3.每二个月清扫冷凝器表面灰尘，冷凝散热不良会引起压缩机部件损坏。清扫时，使用吸尘器、小毛刷等清洗冷凝表面油尘，不能使用尖锐金属工具清扫，以免损坏冷凝器。
4.制冰机的水管、水槽、储冰箱及保护胶片要每两个月清洗一次。不使用时，应清洗干净，并用电吹风吹干冰模及箱内水分，放在无腐蚀气体及通风干燥的地方，避免露天存放。
⑴开机前必须检查自动供水装置是否正常，水箱存水量是否合理（本机出厂时已调正好水位，用户可不作调正）。
⑵插上电源，制冰机开始工作，首先水泵开始运行（水泵有一个短时间的排空气过程）约2分钟后压缩机开始启动，机器进入制冰状态。
⑶当冰块厚度达到设定的厚度时，冰板探针开始启动，除霜电磁阀开始工作，水泵停止工作，热气进入蒸发器，约1分钟左右冰块下落。在冰块下落时，使落冰档板翻转并打开磁簧开关。当磁簧开关重新闭合时，机器进入再一次制冰过程。
⑷压缩机在整个制冰和脱冰过程中都不停机。
⑸当储冰桶内冰满，磁簧开关不能自动闭合时，机器自动停止工作，当取走足够的冰块，磁簧开关重新闭合后，延时3分钟后机器启动，重新进入制冰过程。
2、冰桥厚度的调正
⑴冰桥的厚度应为3mm左右，探针与蒸发器的间际应比实际的冰桥厚度厚1.5mm左右，顺时针旋转调节螺钉口增加冰桥厚度（螺钉旋转1/3圈冰桥厚度改变1.5mm）。
⑵检查冰板探针的接线和连接支架，应保证自由转动使每个制冰过程后都能回到正确的位置。向左转|向右转
谢谢各位！
因为本实验室要建立基因的分离、克隆、表达的平台，需要以下设备，请内行人给个大体的报价，谢谢。我的信箱：zju882003@yahoo.com.cn
此过程如下：PCR扩增目的基因→T-A克隆，酶切鉴定，测序→亚克隆构建表达载体，酶切鉴定，测序→IPTG诱导重组融合蛋白表达→SDS-PAGE检测重组融合蛋白分子量大小→NTA亲和层析法纯化重组融合蛋白→WesternBlot检测融合蛋白免疫反应
1．生化细菌培养箱
2．制冰机
3．PH仪
4．电转化仪（要好的公司提供的产品）
5．蒸馏器
6．高压消毒锅
7．夹心式垂直电泳槽（要好的公司提供的产品）
8．稳压稳流电泳仪
9．磁力搅拌器
10．细菌摇床
11．超声细菌破碎仪
14．转膜仪（要好的产品）
15．融合蛋白的纯化系统（要最好的产品）
16．4℃冰箱
17．微量移液器(1-10ul,10-100ul,100-1000ul)Eppendorf
18．普通照相机
19．紫外分光光度计
20．超净工作台
