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@article{963512,
author = {Masařík, Michal and Gumulec, Jaromír and Sztalmachová, Markéta and Hlavna, Marián and Babela, Petr and Křížková, Soňa and Ryvolová, Markéta and Jurajda, Michal and Sochor, Jiří and Adam, Vojtěch and Kizek, René},
article_number = {24},
doi = {http://dx.doi.org/10.1002/elps.201100301},
keywords = {Immunoprecipitation; Metallothionein; Microparticles; Microseparation; Prostate cancer},
language = {eng},
issn = {0173-0835},
journal = {Electrophoresis},
title = {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},
volume = {32},
year = {2011}
}
TY - JOUR
ID - 963512
AU - Masařík, Michal - Gumulec, Jaromír - Sztalmachová, Markéta - Hlavna, Marián - Babela, Petr - Křížková, Soňa - Ryvolová, Markéta - Jurajda, Michal - Sochor, Jiří - Adam, Vojtěch - Kizek, René
PY - 2011
TI - 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
JF - Electrophoresis
VL - 32
IS - 24
SP - 3576–3588
EP - 3576–3588
SN - 01730835
KW - Immunoprecipitation
KW - Metallothionein
KW - Microparticles
KW - Microseparation
KW - Prostate cancer
N2 - 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 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.
ER -
MASAŘÍK, Michal, Jaromír GUMULEC, Markéta SZTALMACHOVÁ, Marián HLAVNA, Petr BABELA, Soňa KŘÍŽKOVÁ, Markéta RYVOLOVÁ, Michal JURAJDA, Jiří SOCHOR, Vojtěch ADAM a René KIZEK. 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. \textit{Electrophoresis}. roč.~32, č.~24, s.~3576–3588. ISSN~0173-0835. doi:10.1002/elps.201100301. 2011.
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