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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.
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.
