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An immunofluorescent demonstration of nuclear proteins David, Lyle Anacletus

Abstract

This thesis describes the development of an immunofluorescent technique and its application to a cytochemical study of the intracellular distribution of a basic protein component which satisfies several criteria used to identify histone. The technique was developed in three steps: 1. Production of Antibody The method of nuclear protein extraction, the preparation of the inoculum, the immunisation procedure and the species of antigen donor and recipient were all considered contributory of the successful production of antibody. 2. Analyses of Antigen and Antibody Electrophoretic and chromatographic techniques were employed to characterise the nuclear protein antigens. The antiserum was purified by immunological absorption and chromatography. The antigen-antibody responses were studied using electrophoretic and immunodiffusion techniques, both singly and in combination. 3. Application to Cell Preparations Fluorescent antibody provided a protein tracer with which to demonstrate the precise cytological location of analytically defined nuclear proteins. The antibody was produced in the chicken against basic protein from calf thymus nuclei. It was specific for nuclei and chromosomes but was neither organ nor species specific.

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