SUMO2-Rhodamine 110
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.
In stock
SKU
0029
$275.00
Description
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.
Specifications
| Quantity: | 50 μg |
|---|---|
| Molecular Weight: | 11.03 kDa |
| Purity: | >97% by LCMS |
| Readout: | Endpoint/Kinetic |
| Label or Dye: | Rhodamine 110 |
| Detection Method: | Fluorescence |
| Substrate Properties: | Protein-Based Substrate. Typical experimental concentration 50-500 nM. |
| Excitation/Emission (nm): | 485/535 |
| 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
Mass Spectrometry
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
1) Mahajan R., Delphin C., Guan T., Gerace L., Melchior F. "A small ubiquitin-related polypeptide involved in targeting RanGAP1 to nuclear pore complex protein RanBP2." Cell 88:97-107(1997)
2) Kamitani T., Nguyen H.P., Yeh E.T.H. "Preferential modification of nuclear proteins by a novel ubiquitin-like molecule." J. Biol. Chem. 272:14001-14004(1997)
3) Meulmeester E., Kunze M., Hsiao H.H., Urlaub H., Melchior F. "Mechanism and consequences for paralog-specific sumoylation of ubiquitin-specific protease 25." Mol. Cell 30:610-619(2008)
4) Tatham M.H., Geoffroy M.C., Shen L., Plechanovova A., Hattersley N., Jaffray E.G., Palvimo J.J., Hay R.T. "RNF4 is a poly-SUMO-specific E3 ubiquitin ligase required for arsenic-induced PML degradation." Nat. Cell Biol. 10:538-546(2008)
5) Dai X.Q., Kolic J., Marchi P., Sipione S., Macdonald P.E. "SUMOylation regulates Kv2.1 and modulates pancreatic beta-cell excitability." J. Cell Sci. 122:775-779(2009)
6) Cong L., Pakala S.B., Ohshiro K., Li D.Q., Kumar R. "SUMOylation and SUMO-interacting motif (SIM) of metastasis tumor antigen 1 (MTA1) synergistically regulate its transcriptional repressor function."J. Biol. Chem. 286:43793-43808(2011)
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)
2) Kamitani T., Nguyen H.P., Yeh E.T.H. "Preferential modification of nuclear proteins by a novel ubiquitin-like molecule." J. Biol. Chem. 272:14001-14004(1997)
3) Meulmeester E., Kunze M., Hsiao H.H., Urlaub H., Melchior F. "Mechanism and consequences for paralog-specific sumoylation of ubiquitin-specific protease 25." Mol. Cell 30:610-619(2008)
4) Tatham M.H., Geoffroy M.C., Shen L., Plechanovova A., Hattersley N., Jaffray E.G., Palvimo J.J., Hay R.T. "RNF4 is a poly-SUMO-specific E3 ubiquitin ligase required for arsenic-induced PML degradation." Nat. Cell Biol. 10:538-546(2008)
5) Dai X.Q., Kolic J., Marchi P., Sipione S., Macdonald P.E. "SUMOylation regulates Kv2.1 and modulates pancreatic beta-cell excitability." J. Cell Sci. 122:775-779(2009)
6) Cong L., Pakala S.B., Ohshiro K., Li D.Q., Kumar R. "SUMOylation and SUMO-interacting motif (SIM) of metastasis tumor antigen 1 (MTA1) synergistically regulate its transcriptional repressor function."J. Biol. Chem. 286:43793-43808(2011)
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)
