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Ubiquitin activating kit

 
BML-UW0400A-0001 20 tests 167.00 USD
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Replaces Prod. #: BML-UW0400

  • For use with wild type or mutant E2 enzymes in E1 initiated/mediated reactions
  • Contains highly sensitive ubiquitin antibody for detection of ubiquitin and ubiquitin conjugates
For activation of ubiquitin for use in ubiquitinylation experiments. The kit provides the means of generating thioester linked, activated ubiquitin-E1 conjugates, utilizing the first step in the ubiquitin cascade, for investigation of ubiquitin activation, subsequent ubiquitin transfer to/interaction with E2 conjugating enzymes, and their use in the ubiquitinylation of E3 ligases and target substrate proteins. Ubiquitin antibody provided allows high sensitivity detection of ubiquitin and ubiquitin-conjugates.

Product Specification

Application Notes:Uses:
  1. Activation of Ubiquitin for conjugation to wild type or mutant E2 enzymes (user supplied) via thioester bond formation.
  2. Use of Ub-E2 conjugates produced in the subsequent ubiquitin modification of specific target proteins in presence of a dedicated ubiquitin E3 ligase.
  3. Investigation of ubiquitin activation by E1 activating enzyme.
  4. Substitution of the wild type ubiquitin provided with ubiquitin mutants or derivatives (e.g. biotinylated-ubiquitin, Prod. No. BML-UW8705) allowing their activation and subsequent utility in the ubiquitin cascade.
 
Quantity:20 tests
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Kit/Set Contains:20X Ubiquitin Activating Enzyme Solution, Ubiquitin Solution, Ubiquitin Antibody Solution, Ubiquitin protein conjugates rabbit PAb, 20X Mg-ATP Solution, 2X Non-reducing Gel Loading Buffer, 10X Ubiquitinylation Buffer.
 
Scientific Background: The covalent attachment of ubiquitin to proteins (ubiquitinylation) and their subsequent proteasomal degradation plays a fundamental role in the regulation of cellular function through biological events involving cell cycle, differentiation, immune responses, DNA repair, chromatin structure, and apoptosis.
Ubiquitinylation is achieved through three enzymatic steps. In an ATP-dependent process, the ubiquitin activating enzyme (E1) catalyzes the formation of a reactive thioester bond with ubiquitin, in the presence of an Mg2+ cofactor, followed by its subsequent transfer to the active site cysteine of an ubiquitin carrier protein (E2). The specificity of ubiquitin ligation arises from the subsequent association of the E2-ubiquitin thioester with a substrate specific ubiquitin-protein isopeptide ligase (E3), which facilitates the formation of the isopeptide linkage between ubiquitin and its target protein
 
UniProt ID:P0CG47 (UBB), P0CG48 (UBC), P62979 (RPS27A), P62987 (UBA52)
 

Product Literature References

Sumo-dependent substrate targeting of the SUMO protease Ulp1: Z.C. Elmore, et al.; BMC Biol. 9, 74 (2011), Abstract; Full Text

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Ubiquitin, (biotinylated) 

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BML-UW8705-0100 100 µg 160.00 USD
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For Research Use Only. Not for use in diagnostic procedures.
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