Tropomyosin antibody: the specific localization of tropomyosin in nonmuscle cells
An antibody against purified chicken skeletal muscle tropomyosin is used in indirect immunofluorescence to visualize the localization of tropomyosin in a variety of nonmuscle cells. The antibody produces a fluorescent pattern which is very similar to that obtained with an actin-specific antibody. This pattern is composed of fluorescent fibers which are shown to be coincident with the fibers seen with phase-contrast optics. High resolution epifluorescent microscopy reveals that fibers stained with the actin antibody show a continuous fluorescence, while fibers reacted with the tropomyosin antibody show a periodic fluorescence. Measurements indicate that the lengths of the fluorescent segments are variable with an average of 1.2 μm while the spacing between segments is approximately 0.4 μm.
Additional Information© 1975 Rockefeller University Press. Received for publication 14 November 1974, and in revised form 18 February 1975. The author is grateful to Drs. K. Weber and R. Pollack for the generous use of their laboratory facilities. He would also like to thank Drs. J. D. Watson, R. F. Gesteland, M. Howe, R. Goldman, and G. Albrecht- Bühler for their moral support and helpful criticism throughout this work. The author is particularly grateful to Dr. C. Anderson for introducing him to the technique of SDS slab gel electrophoresis and for his invaluable suggestions throughout this work. He also expresses gratitude to Dr. S. Lowey at Brandeis University for her generous gift of purified proteins, to Dr. F. Miller at the State University of New York at Sonty Brook for his continuous advice on the preparation of the antibodies, and to Dr. J. W. Sanger at the University of Pennsylvania Medical School for his critical evaluation of this work. This investigation was supported by a grant from the National Institutes of Health (Research Grant CA 13106 from the National Cancer Institute).
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