K-950

Material Data Sheet

26S Proteasome Degradation Kit

Price: $225.00
Catalog #: K-950

26S Proteasome Degradation Kit

Price: 
$225
Quantity: 
1 Kit

A specific series of specialized chromatographic steps yields a partially purified fraction containing 26S proteasome and none of the conjugation enzymes (E1, E2s, or E3s). This stable fraction is preferred for the degradation of pre-formed ubiquitin-protein conjugates by the 26S proteasome since highly purified 26S is costly and more labile. The 26S concentration is estimated based on analytical SEC at 1500 kDa and quantitation of the 20S core particle activity using a fluorogenic peptide substrate. General procedures for the use of this product are found below.


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

Product Information

Stock: 
1. 26S Fraction [protein] = X mg/ml, 100 μL     Estimated Purity = 60%     Estimated [26S] = X nM in 50 mM HEPES, pH 7.6, 0.5 mM     Mg-ATP, 150 mM NaCl, 10% glycerol2.Energy Solution - 10X Stock, 500 μL
Storage: 

Store solutions at -80°C. Once thawed, keep on ice prior to use. For best results, aliquot and freeze after first set of experiments to avoid multiple freeze/thaw cycles.

Literature

References: 

Ciechanover A. (1998) EMBO. J. 17:7151-7160
Coux O. (1996) Ann. Rev. Biochem. 65:801-847
Hershko A. and Ciechanover A. (1998) Ann. Rev. Biochem. 67:425-479
Ganoth D., et al. (1988) J. Biol. Chem. 263:12412-12419
Glickman M.H and Ciechanover A. (2001) Physiol. Rev. 82:373-428
Schwartz A.L. and Ciechanover A. (1999) Ann. Rev. Med. 50:57-74
Rechsteiner M., et al. (1993) J. Biol. Chem. 268:6065-6068
Voges D., et al. (1999) Ann. Rev. Biochem. 68:1015-1068