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New method of plant mitochondria isolation and sub-fractionation for proteomic analyses
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SYSNO ASEP 0109145 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Ostatní články Název New method of plant mitochondria isolation and sub-fractionation for proteomic analyses Překlad názvu Nová metoda isolace a subfrakcionace rostlinných mitochondrií vhodná pro proteomické analýzy Tvůrce(i) Hájek, Tomáš (UEB-Q)
Honys, David (UEB-Q) RID, ORCID
Čapková, Věra (UEB-Q) RIDZdroj.dok. Plant Science. - : Elsevier - ISSN 0168-9452
Roč. 167, č. 3 (2004), s. 389-395Poč.str. 7 s. Jazyk dok. eng - angličtina Země vyd. IE - Irsko Klíč. slova plant mitochondria isolation ; sub-fractionation ; protein analysis Vědní obor RIV EB - Genetika a molekulární biologie CEP LZ1K03018 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy CEZ AV0Z5038910 - UEB-Q Anotace A new method for the isolation and sub-fractionation of mitochondria from plant tissues is described. This method was used for the analysis of proteins isolated from a pure total mitochondrial fraction and of protein complexes obtained from two mitochondrial sub-fractions, membrane-bound and matrix, from in vitro cultivated tobacco pollen tubes. The method comprised: a new plant tissue homogenization procedure: differential centrifugation of the homogenates in a Percoll gradient; mitochondria immobilization together with Percoll removal by filtration through an uncharged nylon membrane. Mitochondria trapped on the nylon membrane were used for (1) rapid protein extraction; (2) quantitative phenol-chloroform protein extraction; (3) mitochondria sub-fractionation into membrane-bound and matrix fractions. The new method was found to be more efficient than previously published protocols, requiring the use of as little as 180 mg of fresh plant material. It is very fast, reliable, and suitable for proteomic analyses of plant organs and tissue samples with low biomass. Pracoviště Ústav experimentální botaniky Kontakt David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469 Rok sběru 2005
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