Conjugation of the small ubiquitin-like modifier SUMO-1/SMT3C/Sentrin-1 to proteins in vitro is dependent on a heterodimeric E1 (SAE1/SAE2) and an E2 (Ubc9). Although SUMO-2/SMT3A/Sentrin-3 and SUMO-3/SMT3B/Sentrin-2 share 50% sequence identity with SUMO-1, they are functionally distinct. Inspection of the SUMO-2 and SUMO-3 sequences indicates that they both contain the sequence psiKXE, which represents the consensus SUMO modification site. As a consequence SAE1/SAE2 and Ubc9 catalyze the formation of polymeric chains of SUMO-2 and SUMO-3 on protein substrates in vitro, and SUMO-2 chains are detected in vivo. The ability to form polymeric chains is not shared by SUMO-1, and although all SUMO species use the same conjugation machinery, modification by SUMO-1 and SUMO-2/-3 may have distinct functional consequences.

Suitable for use in Western Blot. Other applications not tested.

Western Blot: 1:500-1:1000 observed size: 60 kD positive control: Jurkat or NIH3T3 whole cell lysate Optimal dilutions to be determined by the researcher.

May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Synthetic peptide of human SENP2.

Supplied as a liquid in PBS, 0.2% gelatin, 0.05% sodium azide.

Purified

Covalent attachment of one protein to another is one of the more prominent posttranslational modifications in respect to size and ubiquity. Ubiquitin is the most familiar of the protein modifiers and its activation and transfer to target proteins has been studied extensively. Recently, a new group of ubiquitin-like proteins has been receiving a lot of attention. SUMO, or Sentrin, is one of the most intriguing. SUMO has been implicated in the stabilization of target proteins and/or their localization to sub-cellular complexes.||Conjugation of the small ubiquitin-like modifier SUMO-1/SMT3C/Sentrin-1 to proteins in vitro is dependent on a heterodimeric E1 (SAE1/SAE2) and an E2 (Ubc9). Although SUMO-2/SMT3A/Sentrin-3 and SUMO-3/SMT3B/Sentrin-2 share 50% sequence identity with SUMO-1, they are functionally distinct. Inspection of the SUMO-2 and SUMO-3 sequences indicates that they both contain the sequence psiKXE, which represents the consensus SUMO modification site. As a consequence SAE1/SAE2 and Ubc9 catalyze the formation of polymeric chains of SUMO-2 and SUMO-3 on protein substrates in vitro, and SUMO-2 chains are detected in vivo. The ability to form polymeric chains is not shared by SUMO-1, and although all SUMO species use the same conjugation machinery, modification by SUMO-1 and SUMO-2/-3 may have distinct functional consequences.||Applications: |Suitable for use in Western Blot. Other applications not tested.||Recommended Dilution:|Western Blot: 1:500-1:1000 |observed size: 60 kD |positive control: Jurkat or NIH3T3 whole cell lysate|Optimal dilutions to be determined by the researcher.||Storage and Stability:|May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20°C or colder. Aliquots are stable for at least 24 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.