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
References
Dec 1, 1979·Molecular Immunology·V T Oi, L A Herzenberg
Jul 1, 1979·Hoppe-Seyler's Zeitschrift für physiologische Chemie·W Kufer, H Scheer
Jun 1, 1977·The Biochemical Journal·A S Brown, R F Troxler
Jul 1, 1987·Archives of Biochemistry and Biophysics·T Isono, T Katoh
Jul 1, 1987·Cytometry·D T SasakiE G Engleman
Feb 1, 1988·Photochemistry and Photobiology·K CsatordayD S Berns
Jan 1, 1987·Cytometry·M R LokenK A Kelley
Mar 1, 1987·Cytometry·S AlbertiL A Herzenberg
Aug 1, 1970·Photochemistry and Photobiology·R E Dale, F W Teale
Mar 1, 1983·The Journal of Experimental Medicine·R A ReichertE C Butcher
Nov 1, 1983·Cytometry·H M ShapiroT B Strom
Mar 1, 1984·Cytometry·D R ParksL A Herzenberg
Jun 1, 1982·The Journal of Cell Biology·V T OiL Stryer
Jun 10, 1980·Biochemistry·R MacCollE Traeger
Sep 5, 1980·Biochimica Et Biophysica Acta·B A ZilinskasP C Kahn
Jan 1, 1982·Annual Review of Microbiology·A N Glazer
Jul 7, 1983·Nature·W M GallatinE C Butcher
Jan 1, 1982·Immunological Reviews·R L Coffman
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
Aug 4, 2001·The American Journal of Pathology·A Mikulowska-MennisS A Michie
Aug 16, 2001·Electrophoresis·P J ViskariC L Colyer
May 18, 2001·Analytical Biochemistry·L J McCartneyD J Birch