Human 20S Immunoproteasome Protein, CF

Catalog # Availability Size / Price Qty
E-370-025
Product Details
Citations (8)
FAQs
Reviews (1)

Human 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. 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 Human 20S Immunoproteasome concentration of 0.5-5 nM.
Source
Human peripheral blood mononuclear cell-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-370

Formulation Supplied as a solution in HEPES, NaCl and 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.
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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 multi-complex Immunoproteasome. The structure of the 20S Immunoproteasome is similar to the 20S Proteasome, which is composed of 28 non-identical subunits arranged into four stacked rings (1,2). However, during 20S Immunoproteasome assembly, the three catalytic beta subunits, beta 1, 2, and 5, in the two interior rings of the 20S Proteasome are replaced by three IFN-gamma-inducible catalytic subunits: beta 1i/LMP2, beta 2i/LMP7, and beta 5i/MECL-1 (3). The 20S Immunoproteasome is commonly associated with the 19S, PA28 alpha/beta, or the PA28 gamma regulatory complexes (3,4). 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 (3,5,6). Selective inhibition of 20S Immunoproteasome proteolytic activity using small molecule inhibitors is being examined for therapeutic intervention in cancer and inflammatory diseases (7).  

 This protein has been purified from human peripheral blood mononuclear cells, which have been screened and are negative for hepatitis B surface antigen, antibodies to hepatitis C virus, HIV type 1 antigens, and antibodies to HIV type 1 and 2.

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

Citations for Human 20S Immunoproteasome Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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  1. Discovery of Immunoproteasome Inhibitors Using Large-Scale Covalent Virtual Screening
    Authors: A Scarpino, D Bajusz, M Proj, M Gobec, I Sosi?, S Gobec, GG Ferenczy, GM Keser?
    Molecules, 2019;24(14):.
    Species: Human
    Sample Types: Covalent Compounds
    Applications: Bioassay
  2. Piperlongumine and some of its analogs inhibit selectively the human immunoproteasome over the constitutive proteasome
    Authors: E Bosc, J Nastri, V Lefort, M Valli, F Contiguiba, R Pioli, M Furlan, VDS Bolzani, C El Amri, M Reboud-Rav
    Biochem. Biophys. Res. Commun., 2018;496(3):961-966.
    Species: Human
    Sample Types: Small Molecule
    Applications: Bioassay
  3. Development of a novel immunoproteasome digestion assay for synthetic long peptide vaccine design
    Authors: H Wada, A Shimizu, T Osada, Y Tanaka, S Fukaya, E Sasaki
    PLoS ONE, 2018;13(7):e0199249.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  4. A therapeutic T cell receptor mimic antibody targets tumor-associated PRAME peptide/HLA-I antigens
    Authors: AY Chang, T Dao, RS Gejman, CA Jarvis, A Scott, L Dubrovsky, MD Mathias, T Korontsvit, V Zakhaleva, M Curcio, RC Hendrickso, C Liu, DA Scheinberg
    J. Clin. Invest., 2017;127(7):2705-2718.
    Species: N/A
    Sample Types: Protein
    Applications: Enzyme Assay
  5. Structure of human immunoproteasome with a reversible and noncompetitive inhibitor that selectively inhibits activated lymphocytes
    Authors: RLA Santos, L Bai, PK Singh, N Murakami, H Fan, W Zhan, Y Zhu, X Jiang, K Zhang, JP Assker, CF Nathan, H Li, J Azzi, G Lin
    Nat Commun, 2017;8(1):1692.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  6. Immunoproteasome functions explained by divergence in cleavage specificity and regulation
    Authors: MB Winter, F La Greca, S Arastu-Kap, F Caiazza, P Cimermanci, TJ Buchholz, JL Anderl, M Ravalin, MF Bohn, A Sali, AJ O'Donoghue, CS Craik
    Elife, 2017;6(0):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  7. Clinical activity of carfilzomib correlates with inhibition of multiple proteasome subunits: application of a novel pharmacodynamic assay
    Authors: SJ Lee, K Levitsky, F Parlati, MK Bennett, S Arastu-Kap, L Kellerman, TF Woo, AF Wong, KP Papadopoul, R Niesvizky, AZ Badros, R Vij, S Jagannath, D Siegel, M Wang, GJ Ahmann, CJ Kirk
    Br J Haematol, 2016;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay
  8. Enzymatic discovery of a HER-2/neu epitope that generates cross-reactive T cells.
    Authors: Henle A, Erskine C, Benson L, Clynes R, Knutson K
    J Immunol, 2013;190(1):479-88.
    Species: N/A
    Sample Types: Recombinant Protein
    Applications: Bioassay

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Human 20S Immunoproteasome Protein, CF
By Anonymous on 05/02/2020
Application: Enzymatic activity in vitro

Ordinate: fluorescent count, Em 345/Ex 445
Em band path: 9 nm
Ex band path: 20 nm
Horizontal: time [sec]
Temp: 37 °C
1. i20S(15 nM)/PA28a(150 nM)/S-300(0.01 uM)
2. i20S(15 nM)/PA28a(150 nM)/S-300(0.1 uM)
3. i20S(15 nM)/PA28a(150 nM)/S-300(1 uM)
4. 20S(15 nM)/PA28a(150 nM)/S-300(0.01 uM)
5. 20S(15 nM)/PA28a(150 nM)/S-300(0.1 uM)
6. 20S(15 nM)/PA28a(150 nM)/S-300(1 uM)
The hydrolysis activity of proteasome and immunoproteasome is considered to be lower than that of trypsin. About 15 nM is required in the co-presence of PA28 activator. At that time, the substrate concentration should be about 1 uM for the observation of the saturation level.