UBIQUITINYLATION KIT

For generation of ubiquitin-E2 thioesters for use in ubiquitinylation experiments.

This kit provides the means of generating a range of thioester-linked ubiquitin conjugation enzymes (E2s), utilizing the first two steps in the ubiquitin cascade; for use in the transfer of ubiquitin to E3 ligases; and the subsequent ubiquitinylation of target proteins.

Biotinylated ubiquitin provided supports thioester formation and high sensitivity detection of ubiquitin conjugates.

Sufficient for 50 assays.

Suggested uses:

1. Generation of ubiquitin-E2 thioesters for use in ubiquitinylation experiments.
2. Ubiquitinylation of target proteins in the presence of a dedicated E3 ligase.
3. Activation of ubiquitin for thioester conjugation to novel E2 enzymes.
4. Use of cell lysate or crude fractions/preparations as source of E3 ligases to facilitate ubiquitinylation.
5. Substrate (target) independent in vitro ubiquitinylation reactions.