Use of the Avidin-Biotin Complex for the Localization of Actin and Myosin with Fluorescence Microscopy

A new indirect method for fluorescence localization of proteins making use of the avidin-biotin complex is described. We have prepared both a biotin-modified rabbit heavy meromyosin (BHMM) and a biotin-modified antibody to a smooth muscle myosin. After fixation, cells can be treated with either BHMM...

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Bibliographic Details
Published in:The Journal of cell biology 1977-06, Vol.73 (3), p.783-788
Main Authors: Heggeness, Michael H., Ash, John F.
Format: Article
Language:English
Subjects:
Actins
Actins - isolation & purification
Antibodies
Avidin
Biotin
Brief Notes
Cell Line
Eggs
Fluorescence
Fluorescent Antibody Technique
Gels
Immunoglobulin G
Molecules
Myofibrils
Myosin Subfragments
Myosins - isolation & purification
Reagents
Room temperature
Sodium
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Description
Summary:A new indirect method for fluorescence localization of proteins making use of the avidin-biotin complex is described. We have prepared both a biotin-modified rabbit heavy meromyosin (BHMM) and a biotin-modified antibody to a smooth muscle myosin. After fixation, cells can be treated with either BHMM, which binds to actin, or the biotinyl antibody, which binds to myosin. In a second step the cell are treated with a fluorescent derivative of avidin (Fl-avidin) which binds to the biotinyl proteins and thus indirectly reveals the location of the cellular action or myosin.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.73.3.783