✦ LIBER ✦

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

✍ Scribed by John L. Giannini; Jose Ruiz-Cristin; Donald P. Briskin

Publisher: Elsevier Science
Year: 1988
Tongue: English
Weight: 587 KB
Volume: 174
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(88)90056-5

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✦ Synopsis

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.

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