Recombinant Human His6-Ubiquitin Mutant with K63 only, CF Summary
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins.Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration.The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard.In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
UM-HK630
Formulation | Lyophilized from a solution in HEPES |
Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Reconstitution Calculator
Background: Ubiquitin
Ubiquitin is a 76 amino acid (aa) protein that is ubiquitously expressed in all eukaryotic organisms. Ubiquitin is highly conserved with 96% aa sequence identity shared between human and yeast Ubiquitin, and 100% aa sequence identity shared between human and mouse Ubiquitin (1). In mammals, four Ubiquitin genes encode for two Ubiquitin-ribosomal fusion proteins and two poly-Ubiquitin proteins. Cleavage of the Ubiquitin precursors by deubiquitinating enzymes gives rise to identical Ubiquitin monomers each with a predicted molecular weight of 8.6 kDa. Conjugation of Ubiquitin to target proteins involves the formation of an isopeptide bond between the C-terminal glycine residue of Ubiquitin and a lysine residue in the target protein. This process of conjugation, referred to as ubiquitination or ubiquitylation, is a multi-step process that requires three enzymes: a Ubiquitin-activating (E1) enzyme, a Ubiquitin-conjugating (E2) enzyme, and a Ubiquitin ligase (E3). Ubiquitination is classically recognized as a mechanism to target proteins for degradation and as a result, Ubiquitin was originally named ATP-dependent Proteolysis Factor 1 (APF-1) (2,3). In addition to protein degradation, ubiquitination has been shown to mediate a variety of biological processes such as signal transduction, endocytosis, and post-endocytic sorting (4-7).
This Ubiquitin mutant contains only a single lysine, K63, with all other lysines mutated to arginine. This mutation renders Ubiquitin able to form poly-Ubiquitin chains with other Ubiquitin molecules only via the K63 lysine.
- Sharp, P.M. & W.-H. Li. (1987) Trends Ecol. Evol. 2:328.
- Ciechanover, A. et al. (1980 ) Proc. Natl. Acad. Sci. USA 77:1365.
- Hershko, A. et al. (1980) Proc. Natl. Acad. Sci. USA 77:1783.
- Greene, W. et al. (2012) PLoS Pathog. 8:e1002703.
- Tong, X. et al. (2012) J. Biol. Chem. 287:25280.
- Wei, W. et al. (2004) Nature 428:194.
- Wertz, I.E. et al. (2004) Nature 430:694.
Citations for Recombinant Human His6-Ubiquitin Mutant with K63 only, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products.The data collected includes not only links to publications in PubMed,but also provides information about sample types, species, and experimental conditions.
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- Fine-Tuning of Shh/Gli Signaling Gradient by Non-proteolytic Ubiquitination during Neural PatterningAuthors: P Ma, NN Song, Y Li, Q Zhang, L Zhang, L Zhang, Q Kong, L Ma, X Yang, B Ren, C Li, X Zhao, Y Li, Y Xu, X Gao, YQ Ding, B MaoCell Rep, 2019;28(2):541-553.e4.Species: HumanSample Types: Whole CellsApplications: Bioassay
- Variation in auxin sensing guides AUX/IAA transcriptional repressor ubiquitylation and destructionAuthors: M Winkler, M Niemeyer, A Hellmuth, P Janitza, G Christ, SL Samodelov, V Wilde, P Majovsky, M Trujillo, MD Zurbriggen, W Hoehenwart, M Quint, LIA Calderón VNat Commun, 2017;8(0):15706.Species: N/ASample Types: ProteinApplications: Enzyme Assay
- Proteasome-independent polyubiquitin linkage regulates synapse scaffolding, efficacy, and plasticityAuthors: Q Ma, H Ruan, L Peng, M Zhang, MU Gack, WD YaoProc. Natl. Acad. Sci. U.S.A., 2017;0(0):.Species: HumanSample Types: Cell LysatesApplications: Bioassay
FAQs
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采用6孔板接毒后,铺上低溶解度琼脂糖,第二天就出来了这些病变,正常细胞没有出现这种现象。
按说空斑应该出现多个,但我的六孔板中一个空最多也就出现了2个空斑,也不像文献报道的那样规则,而且很大!
由于没有做梯度稀释,会不会是接毒量过大造成的啊?请大侠指点,万分感谢!
2) 克隆技术使用使倾向于量繁殖现种群利用价值体,按自规律促进整种群优胜劣汰.意义说,克隆技术干扰自进化程.
3) 克隆技术种昂贵技术,需要量金钱物专业士参与,失败率非高.莉277实验唯.虽现发展更先进技术,功率能达2-3%.
4) 转基物提高疾病传染风险.例,产药物牛奶牛染病毒,种病毒能通牛奶染病
5) 克隆技术应用于体导致代遗传性状工控制.克隆技术引起争论核能否允许发育初期类胚胎进行遗传操作.伦理家所能接受.
6) 克隆技术用创造超,或拥健壮体格却智力低.且,克隆技术能够类效运用,男性失遗传意义.
7) 克隆技术家庭关系带影响巨.由父亲DNA克隆孩看作父亲双胞胎兄弟,延迟几十已.难设想,发现自另外完全复制品,(或)受
1.如果在引物的5’引物引入了T7启动子,3‘引物也引入了polyT,PCR扩增出全长片段,回收纯化之后可以用来直接做体外转录么?为什么看到文献里都是先把这个PCR片段连接到载体里面,扩增之后提取出来线性化之后,再进行体外转录。是不是转录对模板的量有要求?
2.如果使用一些带有T7启动子序列的载体比如pBluescriptSK,在启动子序列和酶切位点之间还有多余的一段序列,这段序列也会被启动子转录,这一小段多余的序列是否会对接下来的转染以及病毒拯救带来影响?
3.有人在做反向遗传么,哪些载体可以用来进行体外转录?插入片段为7.5K。
般基克隆物体直接取基包括面几步骤
提取植物或者物基组DNA(试剂盒)————做PCR需要基扩增(顺利2搞定)——扩增基做酶切——基连接想插入载体面(顺利1-2)——鉴定克隆基
步骤都商品化试剂或者试剂盒 要思路清晰做起快 1-2周能完