Identification of a nuclear protein component of interchromatin granules using a monoclonal antibody and immunogold electron microscopy.
A monoclonal antibody, designated 780-3, has been generated which preferentially recognizes an antigenic component of interchromatin granules in human cells. By indirect immunofluorescence procedures, monoclonal antibody 780-3 produces a cell cycle-specific speckled nuclear staining pattern in adult human fibroblasts which is dramatically altered during metaphase. In contrast, transformed cells appear to express this antigen throughout the cell cycle in increased quantities. Immunogold electron microscopy revealed that the nuclear antigen is intimately associated with interchromatin granules in human cells. Analysis by immunoblot procedures showed that monoclonal antibody 780-3 recognizes two polypeptides of 105 and 41 kD. From these data, a possible nucleolar derivation of interchromatin granules is discussed. These studies demonstrate for the first time that monoclonal antibodies may be used in combination with immunogold electron microscopy to identify the ultrastructural location of nuclear antigens.
D000911:Antibodies, Monoclonal; D002467:Cell Nucleus / Q000648:ultrastructure*; D002478:Cells, Cultured; D002843:Chromatin / Q000648:ultrastructure*; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006046:Gold; D006801:Humans; D008938:Mitosis; D008970:Molecular Weight
D000911:Antibodies, Monoclonal; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006046:Gold