Chapter 10 Immunofluorescent Techniques in the Analysis of Chromosomal Proteins

Abstract

Publisher Summary This chapter focuses on immunofluorescent techniques in the analysis of chromosomal proteins. Progress in the analysis of the nonhistone chromosomal proteins (NHC proteins) has been slow because of the complexity of this class of proteins and the fact that many of the major NHC proteins apparently play structural roles for which no direct assay is available. The use of specific antibody techniques appears likely to assist in resolving these problems. Specific antibodies against chromosomal proteins can be used to determine the distribution of these proteins in situ using indirect staining techniques. The in situ distribution patterns obtained can suggest as well as test hypotheses concerning structural and active roles of these proteins. Specific antibodies can also be used for rapid purification of components from complex mixtures and as probes in enzyme assays. The chapter discusses the production of antibodies against chromosomal proteins of Drosophila and their use in an indirect immunofluorescent assay to determine the in situ distribution of these proteins. The chapter discusses the preparation of antigens and antisera and the characterization of antiserum by indirect immunofluorescent staining of sodium dodecyl sulfate (SDS)-polyacrylamide gels.