Material Data Sheet

Fraction II Degradation Kit

Price: $295.00
Catalog #: K-920

Fraction II Degradation Kit

1 Kit

This kit contains reagents to allow for the controlled degradation of exogenously added or endogenously contained substrate proteins through the ubiquitin proteasome pathway (UPP). Fraction II is the protein fraction of a cell extract that binds to anion exchange resin. It contains E1, most E2s, some E3s, UCHs, and the proteasome (20S and 26S); and is essentially free of ubiquitin and ATP.

NOTE: Kit contains reagents sufficient for 10 x 50 μl reactions.

Product Information

1. Fraction II HeLa, 250 μL    [FII] = X mg/ml in 50 mM HEPES, pH 7.6, 1 mM DTT.2. Ubiquitin, 10 X stock, 75 μL    [Ub] = X mg/ml in 50 mM HEPES, pH 7.63. Energy Regeneration Solution (ERS), 10X, 50 μL4. MG-132, Inhibitor Control, 20 μL    [MG-132] = X μM in 100% DMSO
The Ubiquitin Proteasome Pathway (UPP) is the cell’s principle mechanism for protein catabolism. The UPP has been shown to have significant involvement in the regulation of critical cellular processes such as transcription, oncogenesis, cell cycle progression, development, growth, selective elimination of abnormal proteins, and antigen processing.<br>The proteasome is a large, multimeric protease that catalyzes the final step of the UPP intracellular protein degradation. The proteasome exists in multiple forms within the eukaryotic cell, and contained in all isoforms is the catalytic core known as the 20S proteasome. The 20S proteasome (700 kDa) is arranged as four axially stacked heptameric rings with two β-subunit rings sandwiched between two α-subunit rings. The multicatalytic centers are located within the internal cavity of the β-subunits. The 26S Proteasome is defined by having one or two19S regulatory caps on the 20S core particle. It is the 26S that is responsible for the recognition and subsequent degradation of poly-ubiquitinated proteins into smaller polypeptides. This<br>degradation is ubiquitin and ATP dependent and will not be 26S proteasome mediated unless target substrate is first ubiquitinated via the conjugation cascade.

Enzyme/Fraction/Protein Solutions should be stored at -80°C. Avoid multiple freeze/thaw cycles.



Driscoll J. and Goldberg A.L. (1990) J. Biol. Chem. 265:4789-4792
Ganoth D., et al. (1988) J. Biol. Chem. 263:12412-12419
Glickman M.H. and Ciechanover A. (2002) Physiol. Rev. 82:373-428
Hersko A. et al. (1983) J. Biol. Chem. 258:8206-8214
Waxman L., et al. (1987) J. Biol. Chem. 262:2451-2457
Voges D., et al. (1988) Ann. Rev, Biochem. 68:1015-1068