UF-330

Material Data Sheet

Di-Ub (K63-linked) FRET TAMRA Pos3

Price: $269.00
Catalog #: UF-330

Di-Ub (K63-linked) FRET TAMRA Pos3

Price: 
$269
Quantity: 
50 µg

Linkage specific diubiquitin is a substrate for enzymes that cleave the isopeptide linkage between two ubiquitin molecules. These FRET-based DUB substrates with isopeptide bonds between ubiquitin moieties are superior to gel densitometry assays. Since DUBs recognize and cleave substrates with specific steric conditions, each substrate varies in the position of the fluorophore and quencher and must be empirically tested for individual enzymes.

Product Information

Stock: 
 X mg/ml (X μM) in 50 mM Hepes pH 7.5, 150 mM NaCl, 2 mM DTT. Concentration varies with lot number.
MW: 
17 kDa
Purity: 
> 95% by SDS-PAGE

Use & Storage

Use: 

Typical substrate concentrations range from 100-500 nM and typical enzyme concentrations range from 0.5-10 nM but depend on specific assay conditions and method of detection. Recommended assay buffer is 50 mM Hepes pH 7.5, 150 mM NaCl, 2 mM DTT. Fluorescence can be monitored using Ex544 nm and Em572 nm wavelengths.

Storage: 

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

Literature

References: 

Buchberger A., et al. (2002) Trends. Cell. Biol. 12:216-221

Cook W.J., et al. (1992) J. Biol. Chem. 267:16467-16471

Fischer R.D., et al. (2003) J. Biol. Chem. 278:28976-28984

Tenno T., et al. (2004) Genes to Cells. 9:865-875

Varadan R., et al. (2002) J. Mol. Biol. 324:637-647

Varadan R., et al. (2004) J. Biol. Chem. 279:7055-7063