PMID: 2502578Jul 6, 1989Paper

A novel and inexpensive source of allophycocyanin for multicolor flow cytometry

Journal of Immunological Methods
T M Jung, M O Dailey

Abstract

Allophycocyanin (APC) belongs to a family of phycobiliproteins that are well suited as fluorescent reagents for flow cytometric analysis, since they have a broad excitation spectrum, a large Stoke's shift and they fluoresce with a high quantum yield. The widespread use of APC has been limited by the availability of raw material and high cost of the purified phycobiliprotein. We have assessed the suitability of dry, powdered Spirulina platensis, available at health food stores, as an inexpensive source of APC. APC was extracted from Spirulina platensis by overnight treatment with lysozyme, followed by ammonium sulfate precipitation. APC was then separated from phycocyanin (the only other major phycobiliprotein in Spirulina) by elution of bound material from an hydroxylapatite column using an increasing continuous phosphate gradient. APC isolated in this manner retained its normal trimeric structure. The absorbance and fluorescence excitation and emission spectra of the purified phycobiliproteins were identical to those previously shown for C-PC and APC. APC can be stored concentrated at 4 degrees C, frozen at -70 degrees C, or as a saturated ammonium sulfate precipitate, with no subunit dissociation or change in spectral propert...Continue Reading

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Citations

Feb 17, 2005·Analytical and Bioanalytical Chemistry·Christa L ColyerJames P Landers
Nov 3, 2001·Journal of Biotechnology·R Bermejo RománE Molina Grima
Apr 2, 2003·Journal of Biotechnology·K M MinkovaM Ch Busheva
Mar 30, 2000·Journal of Photochemistry and Photobiology. B, Biology·O J RolinskiJ C Pickup
Apr 15, 2010·Avian Pathology : Journal of the W.V.P.A·S De BoeverE Meyer

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