Material Data Sheet

SUMO3 Conjugation Kit

Price: $355.00
Catalog #: K-720

SUMO3 Conjugation Kit

1 Kit
Data Sheet: 

This kit is designed for the conjugation of the Ubiquitin-like modifier SUMO3 to protein substrates in vitro, which requires the activities of the heterodimeric human E1 activating enzyme (SAE1/SAE2) and the Ubc9 E2 enzyme.  This kit does not contain a SUMO E3 ligase, which may be needed effective SUMOylation of some proteins.

NOTE: Kit contains reagents sufficient for 5-10 x 20 μl reactions.

Product Information

Western Blot Data: 
Western Blot Data: 
The Ubiquitin-like SUMO3 is conjugated to a variety of proteins in the presence of Ubc9 and the SUMO E1 activating enzyme (SAE1/SAE2 in human, or Aos1/Uba2p in yeast). This isoform is known to form poly-SUMO chains on substrates via isopeptide bond formation between the C-terminal glycine of one SUMO3 to the lysine-ε-amino group of K11 on the preceding SUMO3. The heterodimeric SAE1/SAE2 complex (38 and 70 kDa respectively) uses ATP to adenylate the C-terminal glycine residue of SUMO3, forming a thioester bond with the SAE2 subunit. The second step is the trans-esterification reaction whereby the activated SUMO3 is transferred to Cys93 of Ubc9. Ubc9 is a member of the E2 family and is homologous to Ubiquitin-conjugating enzymes, but is specific for the conjugation of SUMO to a variety of target proteins. This E2 is unusual in that it sometimes interacts directly with protein substrates that are modified by SUMOylation, and may play a role in substrate recognition. SUMOylated substrates are primarily localized to the nucleus (RanGAP-1, RANBP2, PML, p53, Sp100, HIPK2) but there are also cytosolic substrates(IκBα, GLUT1, GLUT4). SUMO modification has been implicated in functions such as nuclear transport, chromosome segregation and transcriptional regulation, apoptosis and protein function and stability.

Store at -80°C. Avoid multiple freeze/thaw cycles.