Isolation of metallothionein from cells derived from aggressive form of high-grade prostate carcinoma using paramagnetic antibody-modified microbeads off-line coupled with electrochemical and electrophoretic analysis
Jazyk angličtina Země Německo Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
22102136
DOI
10.1002/elps.201100301
Knihovny.cz E-zdroje
- MeSH
- buněčné extrakty chemie MeSH
- časové faktory MeSH
- elektrická impedance MeSH
- elektroforéza v polyakrylamidovém gelu metody MeSH
- fluorescenční mikroskopie MeSH
- imunoglobulin G chemie metabolismus MeSH
- imunoprecipitace metody MeSH
- koncentrace vodíkových iontů MeSH
- lidé MeSH
- lineární modely MeSH
- magnety * MeSH
- metalothionein izolace a purifikace metabolismus MeSH
- mikrosféry MeSH
- monoklonální protilátky chemie MeSH
- nádorové buněčné linie MeSH
- nádory prostaty chemie metabolismus patologie MeSH
- teplota MeSH
- těžké kovy MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- buněčné extrakty MeSH
- imunoglobulin G MeSH
- metalothionein MeSH
- monoklonální protilátky MeSH
- těžké kovy MeSH
Prostate cancer with altered zinc(II) cell metabolism is the second most frequently diagnosed cancer in developed countries. The alterations of zinc(II) metabolism can influence metabolism of other metal ions and can also be associated with the expression and translation of metal-binding proteins including metallothioneins. The aim of this article was to optimize immunoseparation protocol based on paramagnetic beads conjugated with protein G for the isolation of metallothionein. Isolated metallothionein was determined by differential pulse voltammetry Brdicka reaction and SDS-PAGE. Optimal conditions: antigen-binding time - 60 min, temperature - 70°C, and buffer composition and pH - acetate buffer, pH 4.3, were determined. Under the optimized conditions, lysates from 22Rv1 prostate cancer cells treated with various concentrations of cadmium(II) and copper(II) ions were analyzed. We observed strong correlation in all experimental groups and all lysate types (r>0.83 at p<0.041) between metallothionein concentration related to viability and concentration of copper(II) ions and cadmium(II) ions in medium. Moreover, the results were compared with standard sample preparation as heat treatment and SDS-PAGE analysis.
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