Recombinant Human Ubiquitin Mutant K48R Protein, 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-K48R
Formulation | Lyophilized from a solution in deionized water. |
Reconstitution | Reconstitute at10 mg/mL in an aqueous solution. |
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).
Mutation of lysine 48 to arginine renders Ubiquitin (Ub) unable to form poly-Ubiquitin chains via lysine 48 linkages with other Ubiquitin molecules. Ubiquitin K48R can form a Ubiquitin-activating (E1) enzyme-catalyzed active thioester at the C-terminus allowing the molecule to be transferred to the lysines of substrate proteins (monoubiquitination). Ideal for the reduction in poly-Ubiquitin chain length/conjugation rates and determining if poly-Ubiquitin chains are K48 linked.
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Citations for Recombinant Human Ubiquitin Mutant K48R 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.
6Citations: Showing 1 - 6Filter your results:
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- Cbl interacts with multiple E2s in vitro and in cellsAuthors: MS Liyasova, K Ma, D Voeller, PE Ryan, J Chen, RE Klevit, S LipkowitzPLoS ONE, 2019;14(5):e0216967.Species: HumanSample Types: Recombinant ProteinApplications: Ubiquitination
- Stepwise multi-polyubiquitantion of p53 by the E6AP-E6 ubiquitin ligase complexAuthors: Y Masuda, Y Saeki, N Arai, H Kawai, I Kukimoto, K Tanaka, C MasutaniJ. Biol. Chem., 2019;0(0):.Species: HumanSample Types: Recombinant ProteinApplications: Bioassay
- Ubiquitination and regulation of AURKA identifies a hypoxia-independent E3 ligase activity of VHLAuthors: E Hasanov, G Chen, P Chowdhury, J Weldon, Z Ding, E Jonasch, S Sen, CL Walker, R DereOncogene, 2017;0(0):.Species: N/ASample Types: ProteinApplications: Ubiquitination
- Arabidopsis ubiquitin-conjugating enzyme UBC22 is required for female gametophyte development and likely involved in Lys11-linked ubiquitinationAuthors: S Wang, L Cao, H WangJ Exp Bot, 2016;0(0):.Species: Plant - ArabidopsisSample Types: Recombinant ProteinApplications: Enzyme Assay
- Inhibition of a NEDD8 Cascade Restores Restriction of HIV by APOBEC3G.Authors: Stanley D, Bartholomeeusen K, Crosby D, Kim D, Kwon E, Yen L, Cartozo N, Li M, Jager S, Mason-Herr J, Hayashi F, Yokoyama S, Krogan N, Harris R, Peterlin B, Gross JPLoS Pathog, 2012;8(12):e1003085.Species: N/ASample Types: Recombinant ProteinApplications: Bioassay
- The APC/C subunit Mnd2/Apc15 promotes Cdc20 autoubiquitination and spindleassembly checkpoint inactivation.Authors: Foster S, Morgan DMol Cell, 2012;47(6):921-32.Species: N/ASample Types: ProteinApplications: Bioassay
FAQs
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View all Proteins and Enzyme FAQsReconstitution Buffers
Reconstitution Buffer 1 (PBS)
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CD11C:树突细胞,单核细胞,巨噬细胞,中性粒细胞
有几个疑问
1:荧光标记到细胞是标记到细胞表面还是细胞质内?
2:荧光应该随着细胞的分化和增殖逐渐消失?是不是分化增殖越快,荧光消失速度越快?
3:有哪些容易操作,成本便宜的荧光物质?
谢谢各位战友
可以用CCR3的抗体标记其他细胞,再反推中性粒细胞所占的比例吗?肺泡灌洗液中主要有嗜酸性粒细胞,淋巴细胞,中性粒细胞和巨噬细胞。
(2)荧光标记法 : 使用二乙酸荧光素(FDP)、碘化丙啶(PI)或异硫氰酸荧光素钠标记的荧光染料与细胞共孵育,用流式细胞仪检测荧光染色阳性细胞的比率。此法其实是(1)法的“荧光”版,但其在灵敏性和准确性方面明显要优于后者。
(3)硝酸镧(La)示踪法: 在正常的生物组织中镧微粒可沉积于细胞间隙,但不能穿过具有1~ 2nm 微小间隙的细胞膜性结构(包括细胞膜和细胞器膜),也不能穿过细胞间的紧密连接。在膜性结构通透性增高时, 镧微粒则可进入细胞、细胞器和紧密连接内, 并在电镜下显示, 镧盐标记技术被认为是一种有效的监测细胞膜通透性变化的标记技术。
(4)LDH释放法: 在正常情况下,细胞内大分子物质LDH 是不能通过细胞膜的, 但在细胞膜受损伤而通透性增加时,可通过受损的细胞膜释放出来。LDH 能较好地反映细胞膜损伤程度。类似的还有检测细胞外K+的漏出率等。