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Proteasome purification kit

BML-PW1075A

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High integrity kit for purification of proteasome complexes.

Isolation of wild type proteasome complexes from biological samples (cell lysates, tissue extracts, plasma, serum), confirmed by Western blot analysis.
Analysis of captured proteasome complex proteins by proteomic methods, for example to determine complex composition and subunit post-translational modification.
Investigation of variations in proteasome complexes in abnormal cell lines/tissues
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.

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Proteasome purification kit Western blot — 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.

Product Details

Application Notes	For the isolation and enrichment of proteasome complexes.
Contents	Proteasome purification matrix (BML-PW9005-0250) HeLa S100 fraction (Control lysate) (BML-SW8750-0100) Proteasome 20S β5, pAb (BML-PW8895-0010)
Quantity	Provides sufficient material for 10 binding assays.

Handling & Storage

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.
Handling	Avoid freeze/thaw cycles.
Short Term Storage	-20°C
Long Term Storage	-80°C
Shipping	Dry Ice

Regulatory Status	RUO – Research Use Only

Mechanistic Studies of the Multiple Myeloma and Melanoma Cell-Selective Toxicity of the Rpn13-Binding Peptoid KDT-11: Dickson, P., Simanski, S., et al.; Cell Chem. Biol. 27, 1383 (2020), Abstract
Physical and Functional Analysis of the Putative Rpn13 Inhibitor RA190: Dickson, P., Abegg, D., et al.; Cell Chem. Biol. 27, 1371 (2020), Abstract
PTK2 regulates the UPS impairment via p62 phosphorylation in TDP-43 proteinopathy: Lee, S., Jeon, Y., et al.; bioRxiv , (2018)
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
Loss of urokinase receptor sensitizes cells to DNA damage and delays DNA repair: P.B. Narayanaswamy, et al.; PLoS One 9, e101529 (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
Differential effect of HDAC3 on cytoplasmic and nuclear huntingtin aggregates: T. Mano, et al.; PLoS One 9, e111277 (2014), Abstract

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Proteasome purification kit

BML-PW1075A

Product Details

Handling & Storage

Datasheet, Manuals, SDS & CofA

Manuals And Inserts

Certificate of Analysis

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