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Isolation of wild type proteasome complexes from biological samples (cell lysates, tissue extracts, plasma, serum), confirmed by Western blot analysis.
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Analysis of captured proteasome complex proteins by proteomic methods, for example to determine complex composition and subunit post-translational modification.
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Investigation of variations in proteasome complexes in abnormal cell lines/tissues
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Comparison of proteasome complexes isolated from biological samples associated with a particular disease/illness with samples from healthy controls.
This kit provides a gentle and rapid means to purify proteasome complexes from biological samples using an agarose immobilised 20S subunit specific proteasome antibody, without the need for genetic modification of the expressed proteasome. Captured proteasomes are eluted under denaturing conditions prior to analysis by Western blotting or characterisation by proteomic methods.
Western blot analysis of proteasome purification from control lysate. Proteasome purification assay was set-up and run as described in manual. The presence of purified proteasome in starting material, unbound fraction and elution fraction was detected by Western blotting as described in kit manual, using the 20S ‘core’ proteasome antibody provided at a dilution of 1:1000. Key: SM = starting material, UF = unbound fraction and EL = elution fraction.
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Product Details
| Application Notes: | For the isolation and enrichment of proteasome complexes. |
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| Quantity: | Provides sufficient material for 10 binding assays. |
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| Use/Stability: | Upon receipt, the Proteasome Purification Matrix (BML-PW9005) should be stored at 4°C. The Control lysate (BML-SW8750) should be stored at -80°C. Other biochemical components should be stored at -20°C for short term and -80°C for long term. |
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| Handling: | Avoid freeze/thaw cycles. |
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| Shipping: | Dry Ice |
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| Short Term Storage: | -20°C |
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| Long Term Storage: | -80°C |
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| Contents: | Proteasome purification matrix (BML-PW9005-0250)
HeLa S100 fraction (Control lysate) (BML-SW8750-0100)
Proteasome 20S β5, pAb (BML-PW8895-0010) |
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| Regulatory Status: | RUO - Research Use Only |
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Product Literature References
A mammalian nervous-system-specific plasma membrane proteasome complex that modulates neuronal function: K.V. Ramachandran, et al.; Nat. Struct. Mol. Biol.
24, 419 (2017),
Abstract;
Full Text
Interaction of proteasomes and complement C3, assay of antisecretory factor in blood: I. Lönnroth, et al.; J. Immunoassay Immunochem.
37, 43 (2016),
Abstract;
Differential effect of HDAC3 on cytoplasmic and nuclear huntingtin aggregates: T. Mano, et al.; PLoS One
9, e111277 (2014),
Abstract;
Identification of nitric oxide as an endogenous inhibitor of 26 proteasomes in vascular endothelial cells: H. Liu, et al.; PLoS One
9, e98486 (2014),
Abstract;
Loss of urokinase receptor sensitizes cells to DNA damage and delays DNA repair: P.B. Narayanaswamy, et al.; PLoS One
9, e101529 (2014),
Abstract;
General Literature References
Mass spectrometric characterization of the affinity-purified human 26S proteasome complex: X. Wang et al.; Biochemistry 46, 3553 (2007),
An integrated mass spectrometry-based proteomic approach: quantitative analysis of tandem affinity-purified in vivo cross-linked protein complexes (QTAX) to decipher the 26 S proteasome-interacting network: C. Guerrero, et al.; Mol. Cell. Proteomics 5, 366 (2006),
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;
Proteasome plasticity: M. H Glickman & D. Raveh; FEBS Lett. 579, 3214 (2005),
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;
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;
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),
