E2-Ubiquitin Conjugation Kit


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Cat.No.:  EKIT-0458
Product Name:  E2-Ubiquitin Conjugation Kit
Assay Type:  Enzyme activity
Description:  E2-Ubiquitin Conjugation Kit provides the tools necessary for generating a range of thioester-linked ubiquitin-conjugated E2 enzymes for use in ubiquitinylation experiments. Biotinylated ubiquitin (provided in the kit) can be detected with streptavidin-linked enzymes via SDS PAGE and western blotting.This product provides sufficient material for a total of 50 reactions: 4 reactions with each included E2 enzyme.Uses for this kit include:1. Ubiquitinylation of target proteins in presence of dedicated E3 ligase. Panel of E2s provided for generation of E2-Ub thioester conjugates for testing vs. specific E3/target combinations. For example: ubiquitinylation of p53 in the presence of mdm2 (E3) and UbcH5b (E2).2. Activation of Ub for thioester conjugation to novel E2 enzymes (substituted like for like with kit E2s, under directly comparable conditions).
Background:  Ubiquitinylation, the covalent attachment of ubiquitin to proteins, 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 a Mg2+ cofactor, followed by its subsequent transfer to the active site cysteine of a 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.
Warning:  For research use only, not for diagnostic use.

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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