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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 Tyrosinase (TYR)
- 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. RPA238Hu01
Source Prokaryotic expression
Host E.coli
Purity > 95%
UOM 50ug
Predicted Molecular Mass n/a
Predicted isoelectric point n/a
酪氨酸酶(TYR)重组蛋白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
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.
酪氨酸酶(TYR)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
率约0.5-2%。临床上分为节段型白癜风和非节段型白癜风(泛发型白癜风),导致色素缺少的机制目前并不完全清楚。目前有三个主要假说解释白癜风的病理机制:自身免疫、自体细胞毒性/代谢、神经功能紊乱。黑素细胞是来源于神经脊细胞的高度分化的细胞,负责色素合成。黑色素产生的依赖酪氨酸酶相关蛋白家族的参与,包括酪氨酸酶(Tyrosinase)、酪氨酸酶相关蛋白1(TYRP1)和酪氨酸酶相关蛋白1(TYRP1/DCT)等基因。MITF调控的基因主要参与黑素合成、转运和黑素细胞的分化等。我们研究通过慢病毒介导进行
活性激酶:纯化重组蛋白,或经IP富集后保留活性的激酶; · 候选底物:纯化全长蛋白、蛋白片段或合成肽段; · ATP和Kinase Buffer:提供磷酸基团及Mg²⁺等激酶反应所需环境; · 磷酸化检测体系:磷酸化位点抗体、泛p-Ser/Thr/p-Tyr抗体、放射性ATP、Phos-tag或质谱等。 正式实验前先确认重组蛋白纯度和浓度。激酶还要关注活性状态,底物则尽量使用结构完整、可溶性好的蛋白。 如果已经预测出具体磷酸化位点,可以同步准备WT底物和磷酸化位点突变体,例如Ser/Thr常做
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