Recombinant Human 19S Proteasome FLAG-UCHL5 (C88A), CF

Catalog # Availability Size / Price Qty
E-367-025
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Recombinant Human 19S Proteasome FLAG-UCHL5 (C88A), CF Summary

Product Specifications

Purity
>70%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain
Activity
The Human 19S Proteasome is a multi-subunit regulatory component of the 26S proteasome.  Reaction conditions will need to be optimized for each specific application. We recommend an initial Human 19S Proteasome concentration of 0.5-5 nM.
Source
Human embryonic kidney cell, HEK293-derived human 19S Proteasome protein
Predicted Molecular Mass
700 kDa

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

E-367

Formulation

X mg/ml (X μM) in 20 mM HEPES pH 7.5, 20 mM NaCl, 20% Glycerol (v/v), 1 mM DTT

Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Reconstitution Calculator

Reconstitution Calculator

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Background: 19S Proteasome

The 19S Proteasome, also known as PA700, is a multi-subunit regulatory particle that associates with the 20S Proteasome core particle to form the 26S Proteasome. The 19S Proteasome can cap the 20S Proteasome at one or both ends and regulate substrate access to proteolytic activity in an ATP-dependent manner by modulating the conformation of the 20S Proteasome and facilitating substrate access to the 20S Proteasome catalytic core (1-3). The 19S Proteasome recognizes ubiquitinated proteins as well as non-ubiquitinated, misfolded proteins. It catalyzes substrate deubiquination, unfolds substrate proteins, and threads them into the 20S Proteasome catalytic core (4,5). The 19S Proteasome consists of a base subcomplex and a lid subcomplex. The base subcomplex is composed of six AAA+ family members, two scaffolding proteins, and regulatory proteins involved in Ubiquitin recognition (6,7). The 19S Proteasome lid subcomplex contains eight subunits and one deubiquitinating enzyme, POH1/Rpn11 (6,7), present at stoichiometric levels. Two other deubiquitinating enzymes, UCHL5 (also known as Uch37) and USP14 are reversibly associated with the 19S Proteasome (8).

This 19S Proteasome preparation was purified with an inactive UCHL5 enzyme (Cys 88 to Ala mutation) and may be used for deubiquitination of target substrates via POH1 or USP14.

References
  1. Adams, G.M. et al. (1998) Biochemistry 37:12927.
  2. Chu-Ping, M. et al. (1994) J. Biol. Chem. 269:3539.
  3. Kohler, A. et al. (2001) Mol. Cell 7:1143.
  4. Strickland, E. et al. (2000) J. Biol. Chem. 275:5565.
  5. Finley, D. (2009) Annu. Rev. Biochem. 78:477.
  6. Stadtmueller, B.M. & C.P. Hill (2011) Mol. Cell 41:8.
  7. Xie, Y. (2010) J. Mol. Cell Biol. 2:308.
  8. Lee, B.H. et al. (2010) Nature 467:179.
Alternate Names
19S Proteasome; PA700

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