Description
General description
MultiDsk - Ubiquitin-binding protein reagent is a GST fusion protein containing an array of five ubiquitin-associated (UBA) domains from the yeast Ub-binding protein Dsk2 (aa 327-373; UniProt P48510). MultiDsk binds and protects ubiquitinated proteins from deubiquitination. When immobilized on glutathione resins, MultiDsk can be used for affinity pull-down of mono- and poly-ubiquitinated proteins from cell extracts and tissue homogenates. This construct also contains a C-terminal 6xHis sequence and an internal 6xHis sequence between the N-terminal GST and the 5XUBA sequence (PMID 23056298). MultiDsk is expressed in and purified from Rosetta (DE3)pLysS strain of E. coli (Cat. No. 70956).
Product Source: Yeast protein, with a GST tag and a His tag, expressed in e. coli.
Immunogen
Epitope: Ubiquitin
This is an protein reagent, not an antibody as it is a yeast protein, with a GST tag and a His tag, expressed in e. coli.
Application
Affinity Precipitation Assay Analysis: A representative lot of MultiDsk-Ubiquitin-binding protein was immobilized on agarose beads and used to precipitate ubiquitinated proteins from yeast and HEK293 whole cell extracts. (Wilson, D.M., et al. (2012) PLoS One. 7(10):e46398).
Affinity Precipitation Assay Analysis: A representative lot of MultiDsk-Ubiquitin-binding protein was immobilized on agarose beads and used to pull down ubiquitinated proteins from yeast whole cell extracts. (Wilson, D.M., et al. (2013) Cell. 154:983-995).
Biochem/physiol Actions
Mono- and poly-ubiquitinated proteins
Physical form
Format: Purified
Purified ubiquitin-binding protein in buffer containing 25mM Tris-HCl pH7.5, 150mM NaCl, 2mM β-mercaptoethanol with 50% glycerol without preservatives.
Preparation Note
Stable for 1 year at -80°C from date of receipt.
Shipped on Dry Ice. Upon delivery aliquot and store at -80°C. Avoid freeze / thaw cycle
Analysis Note
Evaluated by Affinity Precipitation in yeast whole cell extract.
Affinity Precipitation Assay Analysis: Ubiquitinated proteins were isolated from 1.0 mg yeast whole cell extract by MultiDsk-Ubiquitin-binding protein reagent (7.8 ug to 31.3 ug MultiDsk per pull-down) immobilized on agarose beads
Multiple bands observed.
Other Notes
Concentration: Please refer to lot specific datasheet.
