Monoclonal antibody against microtubule associated protein-1 produces immunofluorescent spots in the nucleus and centrosome of cultured mammalian cells.
A monoclonal antibody was raised against the highest molecular weight protein associated with microtubules (MAP-1). Its specific binding to MAP-1 was determined by immunoblotting of the gel electrophoretogram of microtubule proteins prepared from porcine brain. The antibody reacted only with MAP-1, not with MAP-2, tau or tubulin. Indirect immunofluorescent staining by this antibody showed bright intranuclear spots, the centrosome and the faint meshwork of the cytoplasm in several types of cultured mammalian cells; HeLa, PtK2, human skin fibroblasts, mouse melanoma cells, Chinese hamster ovary cells. The nuclear spots in the interphase cells, were replaced by diffuse enhanced fluorescence throughout the cell except for chromosomes during mitosis. They reappeared in late telophase, first in the cytoplasm, late in the nucleus. The punctate pattern of nuclear immunofluorescence was not affected by microtubule-depolymerizing agents. The result that it persisted on residual cell structures after extraction with a high salt concentration buffer containing Triton X-100 followed by digestion with DNase I and RNase A suggests that the antigen is associated with the nuclear skeleton.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D002467:Cell Nucleus / Q000032:analysis*; D002478:Cells, Cultured / Q000032:analysis / Q000276:immunology; D006224:Cricetinae; D003412:Cricetulus; D003593:Cytoplasm / Q000032:analysis; D005260:Female; D005347:Fibroblasts / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells / Q000276:immunology; D006801:Humans; D008545:Melanoma / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008869:Microtubule-Associated Proteins; D010053:Ovary / Q000166:cytology; D011506:Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D008869:Microtubule-Associated Proteins; D011506:Proteins