R-Phycoerythrin (absorption spectrum 280; 495; sh 535, 564 nm with A564/A280 ratio of 5.3) was purified from the red macroalga Corallina officinalis. The relative molecular mass determined from PAGE was 240000. SDS-PAGE demonstrated two major subunits of Mr 20000 and 21000, respectively, and a minor subunit of Mr 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.Abbreviations: Mr = molecular mass; PAGE = polyacrylamide gel electrophoresis; Rm = relative migration compared to anion front on a gel; SDS = sodium dodecyl sulphate.

No abstract

R-Phycoerythrin (absorption spectrum 280; 495; sh 535, 564 nm with A564/A280 ratio of 5.3) was purified from the red macroalga Corallina officinalis. The relative molecular mass determined from PAGE was 240000. SDS-PAGE demonstrated two major subunits of Mr 20000 and 21000, respectively, and a minor subunit of Mr 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.Abbreviations: Mr = molecular mass; PAGE = polyacrylamide gel electrophoresis; Rm = relative migration compared to anion front on a gel; SDS = sodium dodecyl sulphate.

Summary. Fluorescence polarization measurement during the progress of fluorochromasia has been used to study the response of human lymphocytes to phytohaemagglutinin (PHA) and to tumour-associated antigens, as a basis for the detection of malignant disease.Polarization (P) values of both stimulated and unstimulated lymphocytes decreased with increasing intracellular fluorescence intensity, and with the duration of the fluorochromatic reaction. When these effects were taken into account, there was no significant difference in the change of P following stimulation of lymphocytes from 50 cancer patients or healthy subjects; the magnitude of the response was related more to the age of the donor and to the extent of granulocyte contamination of the lymphocyte preparation than to the presence of cancer.There were, however, significant differences in the change in leakage of fluorescein out of the lymphocytes and in the change in hydrolysis rate after PHA stimulation between lymphocytes from healthy individuals and from patients with cancer.

No abstract