Material Data Sheet
This biotinylated protein can be used as a positive control for sumoylation assays. It is modified
by SUMO-1, SUMO-2 and SUMO-3 proteins and can be detected with avidin-linked reagents.
E2-25K is a class II ubiquitin-conjugating E2 enzyme identified as a target for sumoylation
during a screen for novel SUMO-1 targets. The SUMO modification of E2-25K has been
extensively characterized in vitro (Pichler et al.). This substrate is efficiently sumoylated by the
SUMO activating E1 (SAE1/SAE2 or Aos1/Uba2) and the Ubc9 E2 enzyme in the absence of an
E3 ligase. The E2-25K SUMO acceptor site is unique and does not conform to the yKXE
consensus site, being located at Lys14 and functioning only in the context of an a-helix.
Stock:X mg/ml (X μM) in 50 mM HEPES pH 7.8, 50 mM NaCl, 10% glycerol, 1 mM DTT. Concentration will vary with specific lot number.
Use & Storage
Typical concentration to use in SUMO conjugation experiments in vitro is 1-5 μM depending on conditions.
Store at -80°C. Avoid multiple freeze/thaw cycles.
Purity:> 95% by SDS-PAGE
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