Fluorescence-activated sorting of isolated nuclei: Heterogeneity of histone H5 immunofluorescence in chicken erythrocyte nuclei

• Published in: Experimental cell research Vol. 135; no. 1; pp. 31 - 37
• Main Authors: Mura, Casilda V., Stollar, B.David
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 1981
• Subjects: animal physiology; Animals; Antibodies; Cell Nucleus - analysis; Chickens; Electrophoresis, Polyacrylamide Gel; Erythrocytes - analysis; Fluorescent Antibody Technique; Histones - blood
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/0014-4827(81)90296-2

When Tween-treated isolated chicken erythrocyte nuclei were examined by indirect immunofluorescence with antibodies to histones H5 and H1, heterogeneity in fluorescence was observed. The distribution of nuclei with varying fluorescence intensity was analysed quantitatively with a fluorescence-activated cell sorter. Even with excess anti-H5 antibody, 15–20% of the nuclei did not stain brightly; with anti-H1, more than 90% of the nuclei were bright. Preparative sorting allowed the collection of dim and bright populations separately. Histones were extracted from these populations and analysed by SDS-polyacrylamide gel electrophoresis. The dim nuclei had as much H5 as the bright ones, suggesting that conformational properties or association with other proteins masked the H5 antigenic determinants in the dim nuclei.