A small scale procedure for the isolation of transport competent vesicles from plant tissues

John L. Giannini, Jose Ruiz-Cristin, Donald P. Briskin

Research output: Contribution to journalArticle

Abstract

A microscale method for the isolation of selectively sealed microsomal membrane fractions from plant tissue is presented. The method is based on differential centrifugation in a table top microcentrifuge to accommodate small sample size (10-25 g tissue) and the addition of KI or KCl in the homogenization medium for isolating selectively sealed plasma membrane or tonoplast vesicles. This microscale procedure was found to be useful in isolating membranes from red beet (Beta vulgaris) storage tissue, sugar beet (Beta vulgaris) storage tissue, corn (Zea mays) roots, and soybean (Glycine max) roots. This paper also describes the ability to further purify an enriched red beet plasma membrane fraction on a discontinuous sucrose density gradient, in a microcentrifuge, that is highly competent in ATP-dependent H+-transport. The speed and wide applicability of this procedure make it ideal when a large number of samples need to be processed.

Original languageEnglish (US)
Pages (from-to)561-567
Number of pages7
JournalAnalytical Biochemistry
Volume174
Issue number2
DOIs
StatePublished - Nov 1 1988

    Fingerprint

Keywords

  • H-transporting ATPases
  • membrane transport
  • plasma membrane
  • tonoplast
  • vesicle

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this