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R-phycoerythrin from the macroalgaCorallina officinalis (Rhodophyceae) and application of a derived phycofluor probe for detecting sugar-binding sites on cell membranes
Authors:C M Hilditch  P Balding  R Jenkins  A J Smith  L J Rogers
Institution:1. Department of Biochemistry, University College of Wales, Aberystwyth, Dyfed, SY23 3DD, UK
2. Sigma Chemical Co., Fancy Road, Poole, Dorset, BH17 7NH, UK
3. Department of Immunology, United Medical and Dental Schools of Guy's and St Thomas' Hospitals, The Rayne Institute, St Thomas' Campus, London, SE1 7EH, UK
Abstract:R-Phycoerythrin (absorption spectrum 280; 495;sh 535, 564 nm with A564/A280 ratio of 5.3) was purified from the red macroalgaCorallina officinalis. The relative molecular mass determined from PAGE was 240 000. SDS-PAGE demonstrated two major subunits ofM r 20 000 and 21 000, respectively, and a minor subunit ofM r 30 000. A fucospyranosyl phenylisothiocyanate conjugate was prepared and this novel fluorescent affinity reagent used in conjunction with a flow cytometer to probe fucose-binding sites on blood mononuclear cells. By varying the sugar and using other phycobiliproteins the approach has the potential for simultaneously monitoring different sugar binding sites on subsets of cells within populations.
Keywords:Corallina officinalis  phycoerythrin  fucose  flow cytometry
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