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Merck
CN

14-1121

MultiDsk-Ubiquitin-binding protein reagent

This MultiDsk-Ubiquitin-binding protein reagent is validated for use in Affinity Precipitation Assays for the pull-down of mono- and poly-ubiquitinated proteins.

Synonym(s):

Dsk-Ubiquitin-binding protein reagent, Ubiquitin domain-containing protein DSK2

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.41
eCl@ss:
32160702
Biological source:
Saccharomyces cerevisiae

Key Documents

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biological source

Saccharomyces cerevisiae

technique(s)

affinity precipitation assay: suitable

UniProt accession no.

P48510

shipped in

dry ice

Quality Level

100

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.

Storage Class

10 - Combustible liquids

wgk

WGK 2

flash_point_f

Not applicable

flash_point_c

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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MultiDsk: a ubiquitin-specific affinity resin.
Wilson, MD; Saponaro, M; Leidl, MA; Svejstrup, JQ
Testing null
Proteasome-mediated processing of Def1, a critical step in the cellular response to transcription stress.
Wilson, MD; Harreman, M; Taschner, M; Reid, J; Walker, J; Erdjument-Bromage, H; Tempst et al.
Cell null

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