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- 文献和实验
- 技术资料
- 保存条件:
-20℃
- 保质期:
12个月
- 英文名:
Recombinant Growth Differentiation Factor 9 (GDF9)
- 库存:
100
- 供应商:
上海沪震
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
Organism species Homo sapiens (Human)
Product No. RPA427Hu01
Source Prokaryotic expression
Host E.coli
Purity > 95%
UOM 50ug
Predicted Molecular Mass 17.0kDa
Predicted isoelectric point n/a
Applications SDS-PAGE; WB; ELISA; IP.
生长分化因子9(GDF9)重组蛋白Endotoxin Level <1.0EU per 1µg (determined by the LAL method)
Subcellular Location n/a
Residues Gly320~Arg454 (Accession # O60383) with N-terminal His-Tag
Formulation Supplied as lyophilized form in PBS, pH7.4, containing 5% trehalose, 0.01% sarcosyl.
SEQUENCES
The target protein is fused with N-terminal His-Tag, its sequence is listed below.
MGHHHHHHSG SEF-G QETVSSELKK PLGPASFNLS EYFRQFLLPQ NECELHDFRL SFSQLKWDNW IVAPHRYNPR YCKGDCPRAV GHRYGSPVHT MVQNIIYEKL DSSVPRPSCV PAKYSPLSVL TIEPDGSIAY KEYEDMIATK CTCR
USAGE
Reconstitute in sterile PBS, pH7.2-pH7.4.
STORAGE AND STABILITY
Storage: Avoid repeated freeze/thaw cycles. Store at 2-8oC for one month. Aliquot and store at -80oC for 12 months.
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.
About the MARKER (complimentary)
Effective Size Range: 10kDa to 70kDa.
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.
生长分化因子9(GDF9)重组蛋白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
重组菌的生长温度,降低培养温度是减少包涵体形成的最常用的方法,较低的生长温度降低了无活性聚集体形成的速率和疏水相互作用,从而可减少包涵体的形成[4]。 2、 添加可促进重组蛋白质可溶性表达的生长添加剂,培养E.coli时添加高浓度的多醇类、蔗糖或非代谢糖可以阻止分泌到周质的蛋白质聚集反应,在最适浓度范围内添加这些添加剂不会影响细胞的生长、蛋白质的合成或运输,其它促重组蛋白质可溶性表达的生长添加剂还有乙醇(诱导热休克蛋白的表达)、低分子量的巯基或二硫化合物(影响细胞周质的还原态,从而影响
有差异,批次一致性没有化学成分明确的培养基好; 5)病毒来源的风险和病毒检测成本极高; 6)HPL 来源有限,我国法律对血液制品提取物,包括 HPL 严格控制,无法实现产业化; 因此,HPL 是一个选择,但是仍然不完善,还有很多空间,因此科学家还在努力,寻找更好的办法。 普通 MSC 细胞培养基+重组蛋白因子混合物 是近年来不断优化的一个解决方案,已经无限接近化学成分完全明确了,但是还是有不少的缺点; 1)MSC 细胞生长需要各种蛋白因子,种类较多,20 种左右,这些蛋白和因子形成聚体或颗粒
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