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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃
- 保质期:
12个月
- 英文名:
Recombinant Geranylgeranyl Diphosphate Synthase 1 (GGPS1)
- 库存:
1000
- 供应商:
上海信裕
| Organism species | Homo sapiens (Human) |
| Product No. | |
| Source | Prokaryotic expression |
| Host | E.coli |
| Purity | > 95% |
| UOM | 50ug |
| Predicted Molecular Mass | 37.3kDa |
| Concentration | n/a |
| Applications | SDS-PAGE; WB; ELISA; IP. |
| Endotoxin Level | <1.0EU per 1µg (determined by the LAL method) |
| Residues | Met1~His290 with two N-terminal Tags, His-tag and T7-tag |
| Formulation | Supplied as lyophilized form in PBS, pH7.4, containing 5% trehalose, 0.01% sarcosyl. |
MEKTQETVQR ILLEPYKYLL QLPGKQVRTK LSQAFNHWLK VPEDKLQIII EVTEMLHNAS LLIDDIEDNS KLRRGFPVAH SIYGIPSVIN SANYVYFLGL EKVLTLDHPD AVKLFTRQLL ELHQGQGLDI YWRDNYTCPT EEEYKAMVLQ KTGGLFGLAV GLMQLFSDYK EDLKPLLNTL GLFFQIRDDY ANLHSKEYSE NKSFCEDLTE GKFSFPTIHA IWSRPESTQV QNILRQRTEN IDIKKYCVHY LEDVGSFEYT RNTLKELEAK AYKQIDARGG NPELVALVKH
Stability Test: The thermal stability is described by the loss rate of the target protein. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37oC for 48h, and no obvious degradation and precipitation were observed. (Referring from China Biological Products Standard, which was calculated by the Arrhenius equation.) The loss of this protein is less than 5% within the expiration date under appropriate storage condition.
Protein bands: 10kDa, 14kDa, 18kDa, 22kDa, 26kDa, 33kDa, 44kDa and 70kDa.
Double intensity bands: The 26kDa, 18kDa, 10kDa bands are at double intensity to make location and size approximation of proteins of interest quick and easy.
Ready-to-use: No need to heat, dilute or add reducing agents before use.
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文献和实验牢记N端法则:当N端第二个残基是Leu、Arg、Lys、Phe、Tyr和Trp时,重组蛋白半衰期只有2分钟,而小侧链的氨基酸残基,如Ala,则可以提高蛋白稳定性。因此在设计载体时,要特别注意第二个密码子,避免上述残基的出现。处理高毒性蛋白:毒性极高的重组蛋白,其本底表达就会杀死原核宿主,质粒容易丢失,使其在大肠杆菌中很难得到高表达。采用可表达T7溶菌酶的宿主。T7溶菌酶是T7RNA聚合酶的抑制剂,采用含有T7溶菌酶质粒的宿主,如pLysS,可以降低重组蛋白在大肠杆菌快速生长期的本底
冻融6、ddH2OpH值适当,避免污染如何选择最合适的DNA聚合酶 1、Taq酶——扩增效率最高发现 1969年,美国黄石国家公园温泉 活性 5‘-3’ DNA聚合酶活性 强 5‘-3’ DNA外切酶活性 弱 荧光探针(定量PCR方法) 3‘-5’ DNA外切酶活性 无 错配率每个循环约1/6000 3‘末端加“A” 能 产物可直接进行“TA”克隆 生产质检 诱导大肠杆菌表达、三次柱纯化,分离获得94kD重组蛋白。 无核
的表达系统会造成质粒的不稳定,细胞生长速度的下降和重组蛋白产量的降低[32,33]。Lanzer和Bujard曾对常用的lac启动子-操纵子系统进行了广泛的研究,证明操纵子放在启动子序列的不同位置会造成70倍差异的抑制。将17bp的操纵子置于-10和-35六聚体区之间所形成的抑制比将其放在-35区的上游或-10区的下游要高50-70倍[34]。 启动子的第三个特性是其简便和廉价的可诱导性。大量生产蛋白质最常用的启动子是热诱导(λPL)和化学诱导(trp)启动子。异丙基硫代-β-D-半乳糖
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