Human Ramos Cell 20S Immunoproteasome Protein, CF

Catalog # Availability Size / Price Qty
E-371-025
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Human Ramos Cell 20S Immunoproteasome Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.
Activity

The human 20S Immunoproteasome is able to degrade substrates in an ATP-independent manner in vitro. It can be activated chemically with SDS (0.035%) or by the addition of PA28.  Reaction conditions will need to be optimized for each specific application.  We recommend an initial i20S Proteasome concentration of 0.5­-5 nM.

Source
Human Ramos cell line (RA 1)-derived 20S Immunoproteasome 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-371

Formulation

X mg/ml (X μM) in 50 mM HEPES pH 7.6, 100 mM NaCl, 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

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: 20S Immunoproteasome

The 20S Immunoproteasome is a modified form of the constitutively active 20S Proteasome core particle and is the catalytic subunit of the multi­complex Immunoproteasome. The structure of the 20S Immunoproteasome is similar to the 20S Proteasome, which is composed of 28 non­identical subunits arranged into four stacked rings. However, during 20S Immunoproteasome assembly, the three catalytic beta subunits, beta 1, beta 2, and beta 5, in the two interior rings of the 20S Proteasome are replaced by three IFN gamma -­inducible catalytic subunits: beta 1i/LMP2, beta 2i/MECl­1, and beta 5i/LMP7. The 20S Immunoproteasome is commonly associated with the 19S, PA28 alpha / beta, or the PA28 gamma regulatory complexes. 20S Immunoproteasome expression is enriched in antigen presenting cells of the immune system where the 20S Immunoproteasome selectively degrades intracellular proteins in a manner that optimizes the generation of peptides for MHC class I antigen presentation. Selective inhibition of 20S Immunoproteasome proteolytic activity using small molecule inhibitors is being examined for therapeutic intervention in cancer and inflammatory diseases. This protein complex has been purified from Burkitt's lymphoma cells (Ramos, RA1, CRL-1596) following stimulation with IFN-gamma and IL-2.

References
  1. Cascio, P. et al. (2001) EMBO J. 20: 2357
  2. Ferrington, D.A. & D.S. Gregerson (2012) Prog. Mol. Biol. Transl. Sci. 109: 75
  3. Kim, H.M. et al. (2011) Biochim. Biophys. Acta 1809: 67
  4. Kloetzel, P.M. (2001) Nat. Rev. Mol. Cell Biol. 2: 179
  5. Lee, W. & K.B. Kim (2011) Curr. Top. Med. Chem. 11: 2923
  6. Niewerth, D., et al. (2014) J Hema & Onc. 7:7
  7. Stadtmueller, B.M. & C.P. Hill (2011) Mol. Cell 41: 8
  8. Xie, Y. (2010) J. Mol. Cell Biol. 2: 308
Alternate Names
20S Immunoproteasome

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