Proteasome ELISA kit

The only commercially available kit for proteasome quantification.



BML-PW0575-0001	1 Kit	378.00 USD

This kit provides the means to quantify 20S proteasome concentrations in biological samples using a sandwich ELISA technique, utilizing two 20S proteasome specific antibodies for capture and detection purposes together with a highly sensitive substrate. Sample 20S proteasome levels are determined by comparison to a 20S proteasome calibration curve produced in parallel. This kit provides sufficient material for 1x96 well plate set-up to be run. Suggested uses: 1. Determination of 20S proteasome levels in biological samples (cell lysates, tissue extracts, plasma, serum).

Quantity:	96 wells (24 tests in triplicate)

Kit/Set Contains:	• 1 × 96 well ELISA plate (Prod. No. BML-KW0900) • 1 vial Capture Antibody (Prod. No. BML-KW8100, 25µL)• 1 vial Detection Antibody (Prod. No. BML-KW8155, 25µL)• 1 vial HRP-labelled Goat anti-Rabbit IgG Secondary Antibody (Prod. No. BML-KW0840, 10µL)• 1 vial 20S Proteasome Stock Solution (Prod. No. BML-KW8720, 1mg/mL, 10µL)• 1 bottle 1× Binding Buffer (Prod. No. BML-KW0845, 15mL) • 1 bottle 10× Wash Buffer (Prod. No. BML-KW0850, 75mL) • 1 bottle 1× Blocking Buffer (Prod. No. BML-KW0855, 30mL)• 1 bottle 1× ELISA Buffer (Prod. No. BML-KW0860, 60mL)• 1 bottle 1× Lysis Buffer (Prod. No. BML-KW0865, 20mL) • 1 bottle 1× TMB Substrate Solution (Prod. No. BML-KW0870, 12mL)• 1 bottle 1× Stop Solution (Prod. No. BML-KW0875, 6mL) - Corrosive!! Note: Upon receipt store the full kit at -80°C until required.

Application:	For quantification of 20S proteasome levels in biological samples.

Long Term Storage:	-80°C

Use/Stability:	Defrost all components completely before use. All kit components can be stored for up to 1 week at 4ºC for convenience once defrosted. Stable for up to six months from receipt when stored at -80°C.

Handling:	Avoid freeze/thaw cycles.

Miscellaneous/General:	Proteasomes are non-lysosomal proteolytic complexes localised primarily in the cytoplasm and in the nucleus of eukaryotic cells. The 26S proteasome structure is composed of a 20S proteasome catalytic core complex and one or two 19S regulatory subcomplexes. The 20S core comprises two copies of 14 subunits (7 α–subunits and 7 β–subunits) arranged in a α7β7β7α7 cylindrical array. Varying catalytic subunit composition (β1, β1i; β2, β2i; β5, β5i) results in a variety of possible subtypes. The 19S regulatory subcomplexes, comprised of 6 ATPase and at least 10 non-ATPase subunits, specifically bind ubiquitinylated proteins and provide the 20S core with an ATP-ubiquitin–dependent proteolytic activity. The ubiquitin-proteasome system is the major non-lysosomal system for the degradation of short half-life proteins and peptides that are involved in basic cellular processes, such as cell-cycle regulation and apoptosis, transcriptional regulation, or antigen processing. Thus, protein degradation by the ubiquitin-proteasome pathway has a major regulatory function for proliferation activity and survival of both normal and malignant cells. The 20S proteasome has been detected in normal human blood plasma (known as circulating proteasome), possessing comparatively low specific activity and with a distinct pattern of subtypes. Proteasomes are often overexpressed in cancer cells; abnormally high expression of proteasomes having been found in human leukaemia cells, renal cancer cells and in breast cancer cell lines. In patients suffering from auto-immune diseases, malignant myelo-proliferative syndromes, multiple myeloma, acute and chronic lymphatic leukaemia, solid tumour, sepsis or trauma, the concentration of circulating proteasome has been found to be elevated, to correlate with the disease state, and may have prognostic significance. Proteasome levels have been measured by enzyme-linked immunosorbent assay (ELISA) techniques in cell lysates, serum or plasma samples. This approach has been used to show that proteasome concentrations in peripheral blood are elevated in patients with certain types of malignant diseases, including multiple myeloma, suggesting that circulating proteasome levels may be correlated with tumour burden. The link between elevated circulating proteasome levels and disease activity has also been demonstrated in patients with systemic autoimmune diseases.

Background / Technical Information:	For the Manual please click here.

General Literature References

Extracellular, circulating proteasomes and ubiquitin - incidence and relevance: S.U. Sixt & B. Dahlmann; Biochim. Biophys. Acta 1782, 817 (2008), Abstract;

Immunological methods to quantify and characterize proteasome complexes: development and application: M. Majetschak & L.T. Sorell; J. Immunol. Methods 334, 91 (2008), Abstract;

Circulating proteasome levels are an independent prognostic factor for survival in multiple myeloma: C. Jakob, et al.; Blood 109, 2100 (2007), Abstract;

Protein degradation by the ubiquitin-proteasome pathway in normal and disease states: S.H. Lecker, et al.; J. Am. Soc. Nephrol. 17, 1807 (2006), Abstract;

High plasma proteasome levels are detected in patients with metastatic malignant melanoma: P.E. Stoebner, et al.; Br. J. Dermatol. 152, 948 (2005), Abstract;

Circulating proteasomes are markers of cell damage and immunologic activity in autoimmune diseases: K. Egerer, et al.; J. Rheumatol. 29, 2045 (2002), Abstract;

Role and function of the 26S proteasome in proliferation and apoptosis: C. Naujokat & S. Hoffmann; Lab. Invest. 82, 965 (2002), Abstract;

Antigen processing by the proteasome: P.M. Kloetzel; Nat. Rev. Mol. Cell Biol. 2, 179 (2001), Abstract;

Plasma proteasome level is a potential marker in patients with solid tumors and hemopoietic malignancies: T. Lavabre-Bertrand, et al.; Cancer 92, 2493 (2001), Abstract;

Reconstitution of hybrid proteasomes from purified PA700-20 S complexes and PA28alphabeta activator: ultrastructure and peptidase activities: F. Kopp, et al.; J. Mol. Biol. 313, 465 (2001), Abstract;

Increased prosomal proteins in breast cancer cells and in neighboring normal cells in Parsi and non-Parsi populations: A. Bhui-Kaur, et al.; J. Cancer Res. Clin. Oncol. 124, 117 (1998), Abstract;

The 26S proteasome: subunits and functions: K. Tanaka & C. Tsurumi; Mol. Biol. Rep. 24, 3 (1997), Abstract;

Structure and functions of the 20S and 26S proteasomes: O. Coux, et al.; Annu. Rev. Biochem. 65, 801 (1996), Abstract;

Serum concentration and localization in tumor cells of proteasomes in patients with hematologic malignancy and their pathophysiologic significance: M. Wada, et al.; J. Lab. Clin. Med. 121, 215 (1993), Abstract;

Changes in expressions of proteasome and ubiquitin genes in human renal cancer cells: H. Kanayama, et al.; Cancer Res. 51, 6677 (1991), Abstract;

Abnormally high expression of proteasomes in human leukemic cells: A. Kumatori, et al.; PNAS 87, 7071 (1990), Abstract;