Active Motif,
Tools to analyze nuclear function,
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simple, effective in vitro SUMOylation of proteins with proven controls

Active Motif's SUMOlink™ Kits provide a simple method for generating SUMOylated proteins in vitro. Using the SUMOylation enzyme cascade, carboxy-terminal SUMO-1, SUMO-2 or SUMO-3 peptide substrates are linked covalently to specific lysine residues on the target protein via isopeptide bonds. SUMOlink Kits provide all the reagents required for performing in vitro SUMOylation of target proteins, including positive and negative controls that help ensure success.

Name Format Cat No. Price  
SUMOlink™ SUMO-1 Kit 20 rxns 40120 $475 Buy Now
SUMOlink™ SUMO-2/3 Kit 20 rxns 40220 $475 Buy Now
Figure 1: Specific SUMO-1, SUMO-2 & SUMO-3 labeling of p53.

Western blot analysis of in vitro SUMOylation of p53 with SUMO-1, SUMO-2 and SUMO -3 wild type (WT) isoforms and their corresponding mutated isoforms. A) SUMO-1 analysis: Western Blots incubated with anti-p53 antibody (1:5000 dilution) and anti-SUMO-1 antibody (1 :4000 dilution). B) SUMO-2/3 analysis: Western Blots incubated with anti-p53 antibody (1:5000 dilution) and anti-SUMO-2/3 antibody (1:4000 dilution).

Lane 1: Wild type SUMO protein conjugation reaction.
Lane 2: Mutated SUMO protein conjugation reaction.
Lane 3: No p53 control protein used in the conjugation reaction.

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
  • Human, mouse and rat reactive

Contents & Storage

SUMO-1 or SUMO-2 and SUMO-3 wild-type and mutated proteins, SUMO-1 or SUMO-2/3 antibodies, p53 control protein, p53 antibody, SUMOylation buffer, Protein buffer, E1 activating enzyme and E2 conjugating enzyme. Reagent storage conditions vary from -20°C to -80°C, see manual for details. All reagents are guaranteed stable for 6 months when stored properly.

SUMOlink Kits provide enough reagents to perform 20 in vitro SUMOylations with the WT SUMO protein and 20 in vitro SUMOylations with the Mutated SUMO protein.