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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 Peroxidasin Homolog (PXDN)
- 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. RPM070Hu01
Source Prokaryotic expression
Host E.coli
Purity > 95%
UOM 50ug
Predicted Molecular Mass 23.0kDa
Predicted isoelectric point n/a
Applications SDS-PAGE; WB; ELISA; IP.
Endotoxin Level <1.0EU per 1µg (determined by the LAL method)
过氧蛋白同源物(PXDN)重组蛋白Subcellular Location n/a
Residues Gln1285~Ala1475 (Accession # Q92626) 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.
MGHHHHHHSGSEF-QSDVFR VAEFPHGYGS CDEIPRVDLR VWQDCCEDCR TRGQFNAFSY HFRGRRSLEF SYQEDKPTKK TRPRKIPSVG RQGEHLSNST SAFSTRSDAS GTNDFREFVL EMQKTITDLR TQIKKLESRL STTECVDAGG ESHANNTKWK KDACTICECK DGQVTCFVEA CPPATCAVPV NIPGACCPVC LQKRA
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 过氧蛋白同源物(PXDN)重组蛋白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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文献和实验指南,用户需要确定自己的最佳实验条件以获取最好的成果。 2.1经由加入具有抗原特异性的抗体到适当的孔(1微克/孔)以加入捕获抗体,浓度应为10微克/毫升于涂层缓冲液中,量应为100μL/孔。※捕获抗体应该是纯化的抗体。2.2在4℃下孵育孔盘过夜。2.3加入250μl封闭液到每个孔中。2.4孵育孔盘于室温下1小时。2.5清空孔盘,用PBST(0.05%Tween-20)洗涤孔盘一次。2.6如果待测物为:A,重组蛋白-以稀释剂稀释待测物至100毫微克,30毫微克,10毫微克,3纳克,1纳克
特异性的选择主要需要考虑四个方面:蛋白特异性、种属特异性、实验方法特异性、标记物的特异性。 (1)蛋白特异性 针对需要检测的蛋白查找抗体,几个细节要区分,重组表达的蛋白和内源性蛋白的检测,对抗体的要求是不一样的,注意查看抗体说明书的检测说明。如果重组蛋白不是全长表达,则需要注意抗体的免疫原区域是否在重组蛋白区域内。内源性蛋白最好能清楚其剪切与修饰的方式,特殊表型的蛋白需要进行序列比对,并结合抗体免疫原序列,查看交叉反应的情况。磷酸化蛋白检测需要确定具体位点,不同位点的磷酸化意味
的缓冲液中),但不同的抗原-抗体反应,封闭条件均不相同,没有适合所有系统的封闭液,具体封闭条件(包括封闭液浓度、缓冲液种类、封闭时间、温度等)需自行优化。 注意事项 a. 影响非特异性结合的因素很多,针对不同的免疫原,其封闭条件均可进行优化,没有通用的封闭液,因为每个抗原-抗体反应都具有独特的性质; b. 在选择封闭物时,最重要的标准是信噪比,封闭条件不足,会导致过多的背景噪音,封闭条件过度,会造成信号变弱; c. 可根据不同的检测系统(碱性磷酸酶或辣根过氧
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