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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原位杂交引物要求：
1. 为了避免检测mRNA时基因组序列的干扰,设计探针时尽量选择在跨内含子的部位。
2. 通常探针的长度（既PCR产物长度）以100-400bp为宜，过长不易穿透组织，杂交效率减低；过短特异性低。
而Realtime引物设计：
1.产物长度80-150bp为宜，可延伸到300bp。
而且你做实时定量的时候，还要做标准曲线，确定引物的扩增效率好不好。
CDNASizeFractionation做完后的电泳检测，发现条带的大小都在5000bp左右，感觉是哪里有问题。前面步骤检测很好。不知道有问题没有？请哪位高人指点一下。
我是这样进行预测的：在“SearchMode:”中选择“rRNAscanonly:”，在“sorce”中选择“mito/chloroplast”,粘贴序列，运行程序。得到了21个tRNA，而鱼中应该是22个tRNA，我觉得很奇怪，在“SearchMode:”和“rRNAscanonly:”中进行其它的设置，都不能得到22个tRNA.
我看的一篇文献“CompletemitochondrialDNAsequenceoftheJapanese
flyingfishCypselurushiraii”中也是说22个tRNA,但是我把文章提交的序列（Genbank号为AB182653.）拿到网上提交，只有21个tRNA，我觉得很奇怪。
不知道是怎么回事？请高手指点，那个软件好像是比较权威的。
似乎都是鉴定目的基因是否导入受体细胞 没有鉴定是否成功导入质粒的 鉴定目的基因是否导入受体细胞有四个层次 1 直接鉴定受体细胞中是否有目的基因 用DNA分子杂交 2 鉴定目的基因是否转录 分子杂交（mRNA） 3 目的基因是否表达 抗原-抗体 （蛋白质） 4 看受体细胞发育成的个体是否表现出相关性状
2、人体病灶细胞或者分泌型细胞有载体病毒受体；
3、DNA病毒，且酶切改造后具有复制能力；
4、表达产物对载体病毒没有抑制作用。
不知道是否准确，希望对你有帮助。
