SUMO1 Rhodamine 110
Catalog number: SBB-PS0028, 50 μg
This SUMO1 substrate is C-terminally derivatized with a Glycine-Rhodamine-110 (3,6-diamino-9-(2-carboxyphenyl)) fluorophore. The Gly-Rh110 remains quenched until the amide bond between the C-terminal glycine and the Gly-Rh110 compound is hydrolyzed. The efficiency of quenching combined with the powerful signal upon hydrolysis yields a reagent with unparalleled signal-to-background. SUMO1-Rh110 can be used to study the deSUMOylating activity of hydrolases SENP1 and SENP2, or other deSUMOylating enzymes. The substrate activity of SUMO1-Rhodamine110 was determined by measuring the SENP1 catalyzed release of unquenched Gly-Rh110. The Excitation and Emission of this substrate is 485nm and 535nm respectively. This protein was expressed in E.coli.
Ubiquitin-like protein that can be covalently attached to proteins as a monomer or 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 E3 ligases such as PIAS1-4, RANBP2 or CBX4. 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 SUMO1 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 bis-Gly-Rh110 compound is hydrolyzed to mono-Gly-Rhodamine 110. The efficiency of quenching combined with the powerful signal upon hydrolysis yields a reagent with unparalleled signal-to-background. SUMO1-Rh110 can be used to study the deSUMOylating activity of hydrolases SENP1 and SENP2, or other deSUMOylating enzymes. The substrate activity of SUMO1-Rhodamine110 was determined by measuring the SENP1 catalyzed release of unquenched mono-Gly-Rh110. The Excitatino and Emission of this substrate is 485nm and 535nm respectively.
For Research Use Only, Not For Use In Humans.
|Molecular Weight:||11.5 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 2. LCMS. Analysis of SUMO1-Rhodamine 110 using LCMS intact mass determination indicates purity greater than 97%, and a molecular weight of 11,525 daltons.
Signal : Background
Figure 3. Signal to Background. The signal to background ratio was determined by 100% hydrolysis of 200nM, 100nM, 50nM SUMO1-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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7) Sun X., Li J., Dong F.N., Dong J.T. "Characterization of nuclear localization and SUMOylation of the ATBF1 transcription factor in epithelial cells." PLoS ONE 9:E92746-E92746(2014)