Post-translational modifications alter protein function by modifying activity or intracellular localization. SUMO (small ubiquitin-like modifier) conjugation affects many biological processes including chromosomal organization and function, DNA repair, nuclear transport and signal transduction pathways. Many SUMO substrates are transcription factors, where SUMO modification is often associated with decreased transcriptional activity. However, in some cases SUMOylation increases transcription factor activity. In vertebrates three paralogs, SUMO-1, SUMO-2 and SUMO-3 are expressed.
The SUMOlink method
Combine your protein of interest with the provided E1 and E2 enzymes, SUMO-1 or SUMO-2 & SUMO-3 protein(s) and buffers. After a 3-hour incubation, the reaction is stopped by adding SDS-PAGE Loading Buffer. SUMOylation is then analyzed by Western blot. p53 protein and antibody are provided for use as a positive control, along with a non-functional, mutated isoform of the SUMO protein(s) for use as a negative control. The SUMO-1 or SUMO-2/3 antibody is used to determine the extent to which your target protein and the p53 control protein have been SUMOylated (see Data tab).
- Investigation of SUMOylation effects on transcription factor activity
- Understanding the role of SUMOylation in the regulation of cellular processes
- Identification of novel proteins as targets for SUMO
Why use SUMOlink?
- Simple, effective method for SUMO conjugation and detection of SUMOylated proteins
- Positive control p53 protein and antibody to ensure success
- Complete kit with all required reagents
- Kits for SUMO-1 or SUMO-2/3 are available
- Wild-type and Mutated SUMO proteins are provided
- Use recombinant protein or cell extracts as starting material