Aloe leaves were crushed in a commercial juicer. After removing coarse materials from the juice by centrifugation, solid ammonium sulphate was added to give 40% saturation. The precipitate was collected by centrifugation, dissolved in 0.05 M carbonate-bicarbonate buffer (pH 9.5) and then the solution was centrifuged. Acetic acid (1 M) was added to the supernatant to give a pH of 4.4. After centrifugation, the supernatant and the precipitate were separated. The precipitate was dissolved in 0.05 M phosphate buffer (pH 8.0) and was then chromatographed on a Sephadex G-200 column. A portion of each fraction was tested for detection of hemagglutinating and mitogenic activities and the active fractions were pooled, condensed with a Spectrapor membrane tube (Spectrum Medical Industries, Inc.) and rechromatographed on the same column. The active fraction was called Aloctin A. The acidic supernatant was lyophilized and dissolved in 0.05 M phosphate buffer (pH 8.0) at a suitable concentration and then chromatographed on a Sephadex G-100 column.
Was this article helpful?