TY - JOUR
T1 - Simultaneous localization of myosin and tubulin in human tissue culture cells by double antibody staining
AU - Fujiwara, K.
AU - Pollard, T. D.
PY - 1978
Y1 - 1978
N2 - We have studied the distribution of myosin and tubulin molecules inside the same tissue culture cells by using two antibodies labeled with contrasting fluorochromes. Antimyosin raised against human platelet myosin was labeled with rhodamine. Antitubulin raised against sea urchin vinblastine-induced tubulin crystals was labeled with fluorescein. The two antibodies stained entirely different structures inside the same flat interphase cell: antimyosin bound to stress fibers and antitubulin bound bound to thin, wavy fibers thought to be individual microtubules. Compact interphase cells stained diffusely with both antibodies. From prophase through early anaphase both antibodies stained the mitotic spindle, although the fluorescence contrast between the spindle and the cytoplasm was much higher with antitubulin than with antimyosin. From anaphase through telophase, strong antimyosin staining occurred in the cleavage furrow, while antitubulin stained the region between the separated chromosomes. This study establishes the feasibility of high-resoltion fluorescent antibody localization of pairs of motility proteins in the cytoplasm of single cells, an approach which will make it possible to map out the sites of the various contractile protein interactions in situ.
AB - We have studied the distribution of myosin and tubulin molecules inside the same tissue culture cells by using two antibodies labeled with contrasting fluorochromes. Antimyosin raised against human platelet myosin was labeled with rhodamine. Antitubulin raised against sea urchin vinblastine-induced tubulin crystals was labeled with fluorescein. The two antibodies stained entirely different structures inside the same flat interphase cell: antimyosin bound to stress fibers and antitubulin bound bound to thin, wavy fibers thought to be individual microtubules. Compact interphase cells stained diffusely with both antibodies. From prophase through early anaphase both antibodies stained the mitotic spindle, although the fluorescence contrast between the spindle and the cytoplasm was much higher with antitubulin than with antimyosin. From anaphase through telophase, strong antimyosin staining occurred in the cleavage furrow, while antitubulin stained the region between the separated chromosomes. This study establishes the feasibility of high-resoltion fluorescent antibody localization of pairs of motility proteins in the cytoplasm of single cells, an approach which will make it possible to map out the sites of the various contractile protein interactions in situ.
UR - http://www.scopus.com/inward/record.url?scp=0018189811&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0018189811&partnerID=8YFLogxK
U2 - 10.1083/jcb.77.1.182
DO - 10.1083/jcb.77.1.182
M3 - Article
C2 - 350890
AN - SCOPUS:0018189811
SN - 0021-9525
VL - 77
SP - 182
EP - 195
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 1
ER -