Analysis Note

Multiple bands observed.

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).

General description

Product Source: Yeast protein, with a GST tag and a His tag, expressed in e. coli.

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).

Immunogen

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.

Epitope: Ubiquitin

Other Notes

Concentration: Please refer to lot specific datasheet.

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.

Quality

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

Specificity

Mono- and poly-ubiquitinated proteins

Storage and Stability

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