Recombinant Human Tetra-Ub WT Chains (K48-linked) Biotin, 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.
UCB-210
| Formulation | Lyophilized from a solution in deionized water. |
| Reconstitution | Reconstitute at2 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: Tetra-Ubiquitin
With a predicted molecular weight of 34 kDa, Tetra-Ubiquitin chains are composed of four Ubiquitin monomers that are covalently linked through isopeptide bonds, which typically form between a lysine residue of one Ubiquitin molecule and the C-terminal glycine residue of another Ubiquitin molecule (1). Each human Ubiquitin monomer is 76 amino acids (aa) in length and shares 96% and 100% aa identity with yeast and mouse Ubiquitin, respectively (2). Seven of the 76 aa in Ubiquitin are lysine residues that can participate in poly-Ubiquitin chain formation. Linkage through specific lysine residues is thought to serve as a signal that affects protein degradation, signaling, trafficking, and other cellular processes (3-8).
Linkage specific Tetra-Ubiquitin can also be used to investigate mechanism of binding and recognition by E1 or E2 enzymes, deubiquitinating enzymes, E3 ligases or other proteins that contain Ubiquitin-associated domains or Ubiquitin-interacting motifs (UIMs). This product is formed with wild-type Ubiquitin and linkage-specific enzymes. Tetra-Ubiquitin is the minimal unit necessary for recognition by the 26S proteasome and contains structural characteristic (such as repeating hydrophobic patches) not present in di-Ubiquitin. This product is made with wild-type human recombinant Ubiquitin and linkage-specific enzymes. These chains are modified with biotin via primary amine coupling. This results in multiple biotinylated species modified at the N-terminus, as well as lysine residues. Biotinylated Ubiquitin can be detected using avidin-linked reagents.
- Scheffner, M. et al. (1995) Nature 373:81.
- Sharp, P.M. & W.-H. Li (1987) Trends Ecol. Evol. 2:328.
- Behrends, C. & J.W. Harper (2011) Nat. Struct. Mol. Biol. 18:520.
- Greene, W. et al. (2012) PLoS Pathog. 8:e1002703.
- Henry, A.G. et al. (2012) Dev. Cell 23:519.
- Tong, X. et al. (2012) J. Biol. Chem. 287:25280.
- Wei, W. et al. (2004) Nature 428:194.
- Zhang, J. et al. (2012) J. Biol. Chem. 287:28646.
- Beal R.E., et al. (1995) Biochem. 37:2925-2934.
- Cook W. J., et al. (1992) J. Biol. Chem. 267:16467-16471.
- Cook W. J. et al. (1994) J. Mol. Biol. 236:601-609.
- Phillips C. L., et al. (2001) Acta. Cryst. 57:341-344.
- Piotrowski J. et al. (1997) J. Biol. Chem. 272:23712-23721.
- Tenno T., et al. (2004) Genes to Cells. 9:865-875.
- Wilkinson K.D., et al. (1995) Biochem. 34:14535-14546.
Citation for Recombinant Human Tetra-Ub WT Chains (K48-linked) Biotin, 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.
1Citation: Showing 1 - 1
- Generation and Validation of Intracellular Ubiquitin Variant Inhibitors for USP7 and USP10Authors: W Zhang, MA Sartori, T Makhnevych, KE Federowicz, X Dong, L Liu, S Nim, A Dong, J Yang, Y Li, D Haddad, A Ernst, D Heerding, Y Tong, J Moffat, SS SidhuJ. Mol. Biol., 2017;0(0):.Applications: Bioassay
FAQs
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Reconstitution Buffers
Reconstitution Buffer 1 (PBS)
Staining Reagents
Streptavidin-Alkaline Phosphatase
Streptavidin-Allophycocyanin
NorthernLights Streptavidin NL493
NorthernLights Streptavidin NL557
Avidin-Fluorescein
NorthernLights Streptavidin NL637
Streptavidin-Phycoerythrin
Supplemental Cell Selection Products
MagCellect Streptavidin Ferrofluid
Supplemental ELISA Products
Streptavidin-HRP
EvenCoat Streptavidin Coated Plates
EvenCoat Streptavidin Coated Plates, 5 Pack
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多谢！
不知道能不能直接买到大鼠抗豚鼠血管IgG的ELISA试剂盒。
如果买不到这种试剂盒，用空白板的话（就是反应孔中没有抗原或者抗体包被），把自制的豚鼠血管抗原加到反应孔中进行孵育，能不能使抗原附着在孔壁上？这样的话就可以按照elisa的操作步骤进行大鼠血清检测了。
目前的设想是自制抗原，粉碎豚鼠的血管，制成悬液，通过反复多次的冻融，离心后收集上清液作为抗原（即豚鼠血管抗原）。
然后将抗原加入到elisa的反应孔中（这种是特制的空白板，就是反应孔中没有抗原或抗体包被）进行孵育，使豚鼠血管抗原附着在孔壁上，就是让反应孔充当固相载体，形成固相抗原。倒掉多余的抗原。
再加入待检测的大鼠血清，这样血清中的特异性抗豚鼠血管IgG就可以跟固相的抗原结合，形成固相抗原抗体复合物。
加入酶标的兔抗鼠或者羊抗鼠IgG,形成酶标的抗原抗体复合物。
然后就是一些显色步骤。
现在关键的问题就是抗原加入空白板，能不能形成固相抗原，如果不能跟孔壁附着的话，在后面洗涤的过程中就被洗掉了，那就没办法完成检测了。
请各位大侠给指条明路啊！！！小弟在此多谢了！！！
