Catalog number: SBB-PS0029, 50 μg
This SUMO2 substrate is C-terminally derivatized with a bis-Glycine-Rhodamine-110 fluorophore. The bis-Gly-Rh110 remains quenched until the amide bond between the C-terminal glycine and the rhodamine compound is hydrolyzed. The efficiency of quenching combined with the powerful signal upon hydrolysis yields an unparalleled signal-to-background. SUMO2-Rh110 can be used to study the deSUMOylating activity of hydrolases SENP1 And SENP2, among other deSUMOylating enzymes. The substrate activity of SUMO2-Rhodamine110 was determined by measuring the SENP1 catalyzed release of unquenched Gly-Rh-110. This protein was expressed in E.coli.
SUMO1 is a Ubiquitin-like protein (UBL) that can be covalently attached to proteins as a monomer or as a lysine-linked polymer. Covalent attachment via an isopeptide bond to its substrates requires prior activation by the E1 complex SAE1-SAE2 and linkage to the E2 enzyme UBE2I, and can be promoted by an E3 ligase such as PIAS1-4, RANBP2, CBX4 or ZNF451. This post-translational modification on lysine residues of proteins plays a crucial role in a number of cellular processes such as nuclear transport, DNA replication and repair, mitosis and signal transduction. This SUMO2 substrate is C-terminally derivatized with a bis-Gly-Rhodamine-110 fluorophore. The bis-Gly-Rh110 is quenched until the amide bond between the C-terminal glycine and the rhodamine compound is hydrolyzed. The efficiency of quenching combined with the powerful signal upon hydrolysis yields an unparalleled signal-to-background. SUMO2-Rh110 can be used to study the deSUMOylating activity of hydrolases SENP1 and SENP2, among other deSUMOylating enzymes. The substrate activity of SUMO2-Rhodamine110 was determined by measuring the SENP1 catalyzed release of unquenched Gly-Rh-110. The Excitation and Emission of this substrate is 485nm and 535nM respectively.
For Research Use Only, Not For Use In Humans.
|Molecular Weight:||11.03 kDa|
|Purity:||>97% by LCMS|
|Label or Dye:||Rhodamine 110|
|Substrate Properties:||Protein-Based Substrate. Typical experimental concentration 50-500 nM.|
|Storage Buffer:||50mM Hepes pH 7.5, 100mM NaCl|
|Storage||Store at −80°C after product arrival. Avoid multiple freeze / thaws. It is recommended to make multiple aliquots after the first thaw.|
Figures & Data
Figure 1. LCMS. Analysis of SUMO2-Rhodamine 110 using LCMS intact mass determination indicates purity greater than 97%, and a molecular weight of 11,030 daltons.
Signal : Background
Figure 2. Signal to Background. The signal to background ratio was determined by 100% hydrolysis of 200nM, 100nM, 50nM SUMO2-Rhodamine 110 to liberate the quenched conjugate. Assay Buffer: 50mM HEPES pH7.5, 1mM TCEP, 0.1mg/ml BSA.
Certificates of Analysis (COA)
Citations & References
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