AQUApure Linear Di-Ub Chains (M1-linked) Protein, CF Summary
Ubiquitin chains vary in length, linkage, and function. Linear (Met1)-linked Di-Ubiquitin Chains (Ub2) are ideal for investigating Ubiquitin-binding proteins and as substrates for Ubiquitin-specific isopeptidases. Reaction conditions will need to be optimized for each specific application. IMPORTANT: Heating this product in SDS-PAGE buffer or terminating reactions containing this product with heated SDS-PAGE buffer could lead to unexpected, high apparent molecular weight banding or smearing on gels that is not representative of product purity. For optimal results, we recommend incubation in SDS-PAGE buffer + DTT at <40 °c="" for="" 20="" minutes="" prior="" to="" gel="">
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.
UC-700B
| Formulation | 1 mg/ml (58 μM) in sterile, deionized water |
| Shipping | The product is shipped with polar packs. 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: Di-Ubiquitin
Linkage specific Poly-Ubiquitin chains may be used as a substrate for in vitro reactions with deubiquitinating enzymes ("DUB"s") that cleave the peptide or isopeptide linkage between adjacent Ubiquitin molecules. Poly-Ubiquitin chains can also be used to investigate mechanisms of binding and recognition between the chains and other proteins that contain Ubiquitin-Associated domains (UBAs), Ubiquitin-interacting motifs (UIMs), ZnF"s and/or other Ubiquitin-sensing elements.
Linear ("M1")-linked Di-Ubiquitin chains are manufactured using recombinant methods to avoid the potential for contaminating synthetic intermediates. The correctness of linkage and purity of each production lot is assessed using the Absolute Quantitation of Ubiquitin method (Ub-AQUA), an LCMS-based technique that provides extremely accurate information on the composition of Poly-Ubiquitin samples.
- Kirkpatrick D.S., et al. (2006) Nat Cell Biol. 8(7) : 700-10
- Ordureau, A., et al. (2014) Mol. Cell 56(3) : 360–375
- Ordureau, A., et al. (2015) Pro. Nat. Acad. of Sci. USA 112(21) : 6637–6642
- Phu L., et al. (2011) Mol Cell Proteomics 10(5) : M110.003756
Citations for AQUApure Linear Di-Ub Chains (M1-linked) Protein, 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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- Post-translational Modification of OTULIN Regulates Ubiquitin Dynamics and Cell DeathAuthors: T Douglas, M SalehCell Rep, 2019;29(11):3652-3663.e5.Species: HumanSample Types: ProteinApplications: Bioassay
- ZUFSP Deubiquitylates K63-Linked Polyubiquitin Chains to Promote Genome StabilityAuthors: P Haahr, N Borgermann, X Guo, D Typas, D Achuthanku, S Hoffmann, R Shearer, TK Sixma, N MailandMol. Cell, 2018;0(0):.Species: HumanSample Types: Cell LysatesApplications: Bioassay
- Cholesterol and fatty acids regulate cysteine ubiquitylation of ACAT2 through competitive oxidationAuthors: YJ Wang, Y Bian, J Luo, M Lu, Y Xiong, SY Guo, HY Yin, X Lin, Q Li, CCY Chang, TY Chang, BL Li, BL SongNat. Cell Biol., 2017;19(7):808-819.Species: N/ASample Types: Applications: Control
- Myosin VI Contains a Compact Structural Motif that Binds to Ubiquitin ChainsAuthors: F He, HP Wollscheid, U Nowicka, M Biancospin, E Valentini, A Ehlinger, F Acconcia, E Magistrati, S Polo, KJ WaltersCell Rep, 2016;14(11):2683-94.Species: HumanSample Types: Recombinant ProteinApplications: Bioassay
- USP45 deubiquitylase controls ERCC1-XPF endonuclease-mediated DNA damageresponses.Authors: Perez-Oliva A, Lachaud C, Szyniarowski P, Munoz I, Macartney T, Hickson I, Rouse J, Alessi DEMBO J, 2015;34(3):326-43.
- Deubiquitinase-based analysis of ubiquitin chain architecture using Ubiquitin Chain Restriction (UbiCRest).Authors: Hospenthal, Manuela, Mevissen, Tycho E, Komander, DavidNat Protoc, 2015;10(2):349-61.Species: HumanSample Types: Protein
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完整的表达载体必须包括：
1、复制子,在细菌中扩增时所必须.
2、启动子,目的基因在细菌或细胞中转录所必须,转录了才能翻译,是谓“表达”.
3、原核筛选标记,细菌增菌所必须.
4、真核筛选标记,如果是真核表达,就是必须的.
5、多克隆位点,即酶切位点,插入目的片段的区域.
6、其他所需构件,可视实验设计情况而选用现成载体或在载体上自行添加.
另：表达的起始和终止,在目的基因上附带起始密码子和终止密码子.
小胶质细胞(microglia)是神经胶质细胞的一种，相当于脑和脊髓中的巨噬细胞，是中枢神经系统(CNS)中的第一道也是最主要的一道免疫防线。小胶质细胞大约占大脑中的神经胶质细胞的20%。小胶质细胞不停地清除着中枢神经系统中的损坏的神经，斑块及感染性物质。无数临床上和神经病理学研究表明激活的小胶质细胞在神经退化类疾病的发病机理中起到十分重要的作用，如帕金森病，多发性硬化和阿兹海默症等。但是过多激活或失控的小胶质细胞会引起神经毒性。他们是促炎因子和氧化应激的重要来源，如肿瘤坏死因子（TNF），一氧化氮，白介素等有神经毒性的物质
exosome，外泌体，是由细胞分泌到外周体液进行细胞间以及细胞与远程组织、器官之间进行信息交流的一种媒介。当前逐渐成为国际研究热点。目前国内刚刚起步，虽然研究成果较国外研究机构尚有不少差距，但已有迎头赶上之势。
外泌体只是胞外囊泡的一种，目前尚未有明确定论胞外囊泡中其它成分的功能。
为了跟随时代科学研究进步的步伐，我建议开通“胞外囊泡及外泌体”讨论区，以促进该领域的充分讨论和合作，弥补当前该领域讨论内容分散，资料不容易汇集等缺点。同时也为广大胞外囊泡及外泌体医学研究者提供一个更好的交流空间。
