Structural,immunocytochemical and initial biochemical characterization of NAOH-extracted gap junctions from an insect,Heliothis virescens |
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Authors: | J. S. Ryerse |
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Affiliation: | (1) Department of Pathology, St. Louis University School of Medicine, 1402 South Grand Avenue, 63104 St. Louis, MO, USA |
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Abstract: | Subcellular fractions enriched in gap junctions with an ultrastructure similar to those in intact insect tissue have been obtained by extracting crude membranes from the tobacco budworm Heliothis virescens (Lepidoptera: Noctuidae) with 2.5 mM NaOH. n-Octyl--d-glucopyranoside (OG) was used to further purify integral membrane proteins in the NaOH-extracted fractions. A polyclonal antibody (R16) is described that specifically labels nonextracted and NaOH-extracted gap junctions in cell fractions by electron microscope immunocytochemistry. R16 immunostaining of sectioned Heliothis testis at the light-microscope level yields a pattern of immunoreactivity consistent with the distribution of gap junctions in the tissue. R16 identifies a 40-kDa protein as a candidate gap junction protein on immunoblots of crude membrane, NaOH-extracted and NaOH/OG-extracted fractions. |
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Keywords: | Gap junction Cell junction Immunocytochemistry Biochemistry Heliothis virescens (Insecta) |
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