SI3304B: Biotinylated K33-Linked Tetra-Ubiquitin

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Specifications

Biotinylated K33-linked ubiquitination is traditionally associated with regulation of the innate immune response. These chain types are also involved in protein stabilization and other non-degradative processes. K33 Tetra-Ubiquitin is a tetrameric chain of wild-type ubiquitin, wherein ubiquitin monomers are enzymatically linked together via an isopeptide bond between Lysine 33 and the C-terminal Glycine. These chain types are also involved in protein stabilization and other non-degradative processes. It is a useful substrate for identifying and characterizing deubiquitinating enzymes that cleave the K33-linkage and for structural and binding studies of ubiquitin chain recognition by ubiquitin-associated domains (UBA) or ubiquitin-interacting motifs (UIMs). Once the K33 Tetra Ubiquitin has been formed we then biotinylate it once on one available site on the ubiquitin chain. This biotin then acts as a means of detection via streptavidin or vidin. The applications are numerous from Western Blots to ELISAs. Testing can vary based off of intended application (EX: DUB activity/specificity).

Info

Species Human
Source E. coli
Tag Biotin
Molecular Weight 35207 Da
Quantity Variable
Concentration Variable
Formulation 20 mM Tris pH 7.5, 150 mM NaCl
Storage -80°C, avoid freeze/thaw cycles

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  • Biotinylated chains are meant to label the protein so they can be selectively separated via a multitude of methods such as Western Blotting, pull downs (see website), ELISAs, etc.
  • Can use streptavidin or avidin to visualize this protein specifically.
  • Investigation of ubiquitin chain specificity and selectivity
  • Studies on the role of ubiquitin chains in protein degradation pathways (e.g., proteasomal and autophagic degradation)
  • Analysis of ubiquitin-mediated signaling pathways and cellular responses
  • Structural studies to elucidate the architecture and dynamics of ubiquitin chains
  • Screening assays to identify modulators of ubiquitin chain assembly and disassembly processes