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- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 保存条件:
-20℃
- 保质期:
12个月
- 英文名:
Recombinant Glutamate Cysteine Ligase, Catalytic (GCLC)
- 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 Mus musculus (Mouse)
Product No. RPD759Mu01
Source Prokaryotic expression
Host E.coli
Purity > 95%
UOM 50ug
Predicted Molecular Mass 24.2kDa
Predicted isoelectric point n/a
Applications SDS-PAGE; WB; ELISA; IP.
Endotoxin Level <1.0EU per 1µg (determined by the LAL method)
谷氨酸半胱氨酸连接酶催化亚基(GCLC)重组蛋白
Subcellular Location n/a
Residues Asp435~Ser636 (Accession # P97494) 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-DFENSA YVVFVVLLTR VILSYKLDFL IPLSKVDENM KVAQKRDAVL QGMFYFRKDI CKGGNAVVDG CSKAQSSSEP AAEEYTLMSI DTIINGKEGV FPGLIPILNS YLENMEVDVD TRCSILNYLK LIKKRASGEL MTVARWMREF IANHPDYKQD SVITDEINYS LIWKCNQIAD ELCECPELLG SGFRKAKYSG GKSDPS
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.
谷氨酸半胱氨酸连接酶催化亚基(GCLC)重组蛋白Ready-to-use: No need to heat, dilute or add reducing agents before use.
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文献和实验基酸有利于甲硫氨酸氨肽酶催化的N-末端甲硫氨酸的去除,从而暴露出位于第二位的亮氨酸,使得该蛋白不稳定[183]。蛋白质的第二个决定因素是位于近氨基端的特异性内源赖氨酸残基[142,143]。该残基是多遍在蛋白链的受体,多遍在蛋白链在真核细胞中有利于遍在蛋白依赖的蛋白酶对蛋白质的降解。有趣的是在一个多遍在蛋白中,它的两个决定簇可以位于不同的亚基上,却能靶向同一个蛋白进行加工[184]。氨基酸成分和蛋白质不稳定性的另一个关系体现在PEST假说中[185]。根据对短寿命真核蛋白的统计分析,蛋白质如果富含Pro
-------------------------- jasperxu (感谢jasperxu战友!) pelB 序列是自己加的?换一个载体试试。我用XL1-Blue preparing M13, it is fine. I use pComb. ------------------------------- yxiangmind (感谢yxiangmind战友长期的帮忙) 在ecoli中用pelB信号肽引导分泌,发现表达量比单独表达高很多?能否详细说明一下。所谓的单独表达是否为胞内非融合表达即ATG直接和目的基因连接? 细胞
是没有问题的. 2、表达重组蛋白时,细菌裂解方法都有哪些? 在表达重组蛋白时,诱导以后跑SDS-PAGE发现表达都很好,但是在裂解细胞时遇到问题。总是不能彻底裂解细胞。 首先我采用超声裂解,可是不管我如何超声总有部分细菌没有裂解。 我的超声条件如下:宁波新芝超声细胞破碎仪,功率400W,超5秒,间隔5秒,重复99次。然后显微镜观察,不彻底时再重复99次。菌体来自200 ml培养液,用20 ml PBS重悬。 然后我又尝试加溶菌酶,首先加至终浓度100 ug/ml,4C半小时菌液不变粘
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