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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 Phosphoinositide Dependent Protein Kinase 1 (PDPK1)
- 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. RPC246Mu01
Source Prokaryotic expression
Host E.coli
Purity > 95%
UOM 50ug
Predicted Molecular Mass n/a
Predicted isoelectric point n/a
Applications SDS-PAGE; WB; ELISA; IP.
Endotoxin Level <1.0EU per 1µg (determined by the LAL method)
Subcellular Location n/a
Residues n/a
Formulation Supplied as lyophilized form in PBS, pH7.4, containing 5% trehalose, 0.01% sarcosyl.
USAGE
Reconstitute in sterile PBS, pH7.2-pH7.4.
STORAGE AND STABILITY
磷酸肌醇依赖性蛋白激酶1(PDPK1)重组蛋白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.
磷酸肌醇依赖性蛋白激酶1(PDPK1)重组蛋白Ready-to-use: No need to heat, dilute or add reducing agents before use.
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文献和实验重组蛋白纯化基本原则蛋白的分离纯化是生物工程下游阶段一个比较重要的部分,尤其在基因工程重组蛋白的分离纯化中,上游过程的许多因素会直接影响到下游蛋白的分离,充分利用上游对下游的影响,对蛋白的纯化做一个全面的考虑和整体的设计。是否带有亲和标签,His标签,GST标签等,不同的亲和标签选择不同的纯化方案;可能是可溶性表达,可能形成包涵体,可溶性的蛋白往往需要复杂的纯化步骤,而包涵体易于分离,纯度较高,但回收具有生物活性的蛋白却变的相当困难,需要对聚集的蛋白进行变复性,通常活性蛋白的得率比较低;是否
,我们用已知的人工合成 MAP 作为阳性对照,分析研究拟南芥的不同 MAP 激酶 [23] 。 ( 4 ) 重组蛋白质除了含有各自的蛋白质的编码序列外,常常还包含由克隆载体所编码的 3' 或 5' 标记物。这些标记物的磷酸化可能会导致假阳性结果,因此在芯片实验之前要将其排除掉。用一些已知的不被认为是相应激酶底物,并且具有相同表达模式(相同标记物)的重组蛋白质,在溶液中进行活性激酶的检测实验可能是一种解决问题的办法。在这个检测实验中,这些所选择的蛋白质应该显示阴性
、Glu、Ser和Thr的区域,且在该区域附近有某些特定的氨基酸,则该蛋白就会不稳定。这些PEST结构域的磷酸化导致钙的结合能力提高,从而利于钙依赖性蛋白酶对蛋白质的降解。这提示可以在缺乏PEST蛋白裂解系统的E.coli中表达PEST富含蛋白。 减少E.coli中重组蛋白裂解的策略有以下几种:(1)将蛋白质靶向细胞周质或培养基[145,186];(2)在较低的温度下培养细菌[187];(3)选用蛋白酶缺陷的菌株[188];(4)构建N-末端或C-末端融合蛋白[186];(5)将目的基因多拷贝串联
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