Proteasome 26S degradation kit



BML-PW8950-0001	1 Kit	241.00 USD

Replaces Prod. #: ALX-850-231

This kit contains a highly purified, human erythrocyte derived, preparation of ‘26S’ proteasomes useful for carrying out in vitro protein degradation studies with suitably ubiquitinylated protein substrates. The preparation consists of a high purity mixture of ‘26S’ proteasomes singly (26S) and doubly (30S) capped with 19S regulatory subunit complexes in the ratio of 40% single cap : 60% double capped at the time of preparation. Additional kit components include ATP for proteasomal activation.

Product Specification

Quantity:	96 assays

Kit/Set Contains:	MgATP (Prod. No. BML-EW9805)(2mM)26S Proteasome (Prod. No. BML-PW9310)(5-10nM; in TSD Buffer (10mM TRIS, 25mM potassium chloride, 1.1mM magnesium chloride, 0.1mM ethylenediaminetetraacetic acid, 1mM dithiothreitol, 1mM sodium azide, 2mM ATP, pH 7.0), 35% glycerol)Radio-labelled ubiquitinylated protein conjugate [NOT SUPPLIED]

Long Term Storage:	-80°C

Use/Stability:	Care must be exercised in the handling and storage of the 26S proteasome to ensure that its activity is not comprimised. The complex should be stored at -80°C. When ready for use it shold be thawed by standing on ice (i.e. not thawed rapidly!). For those wishing to measure and use the enzymatic activity of 26S proteasome it should be used immediately it is thawed since the enzyme complex is labile after thawing After dissociation of the complex, the 20S proteasome activity is relatively stable.

Background / Technical Information:	Please click here for the comprehensive product datasheet.

Product Literature References

Reconstitution of hybrid proteasomes from purified PA700-20 S complexes and PA28alphabeta activator: ultrastructure and peptidase activities: F. Kopp et al.; J. Mol. Biol. 313, 465 (2001), Abstract;

A multicomponent system that degrades proteins conjugated to ubiquitin. Resolution of factors and evidence for ATP-dependent complex formation: D. Ganoth et al.; J. Biol. Chem. 263, 12412 (1988), Abstract;

The protein substrate binding site of the ubiquitin-protein ligase system: A. Hershko et al.; J. Biol. Chem. 261, 11992 (1986), Abstract;

ATP-dependent degradation of ubiquitin-protein conjugates: A. Hershko et al.; PNAS 81, 1619 (1984), Abstract;

Components of ubiquitin-protein ligase system. Resolution, affinity purification, and role in protein breakdown: A. Hershko et al.; J. Biol. Chem. 258, 8206 (1983), Abstract;