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Isolation of plasma membranes from Drosophila embryos

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Abstract

Cell surface components probably play an important role in early embryonic development. However, hardly any information is available on the structure or regulation of expression of the corresponding genes. As a first step in approaching this issue, we devised a procedure to obtain enriched plasma membranes from embryonic Drosophila cells. Membranes are fractionated according to two independent physical parameters: size, using velocity gradient centrifugation and density, using isopicnic gradient centrifugation. The final membrane fraction is enriched by 6 to 8 fold with respect to the plasma membrane enzyme marker Na+/K+ ATPase and substantially depleted of the mitochondrial enzyme marker cytochrome C oxidase. Two-dimensional polyacrylamide gel electrophoresis of the purified membranes reveals enrichment for specific proteins and electron microscopy reveals membrane vesicles in abundance. The enriched fraction should be suitable for the preparation of antibody probes that recognize cell surface components.

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Author notes

  1. Quiao-Yun Jiang

    Present address: Department of Biology, University of Science and Technology of China, Hefei, Anhui, People's Republic of China

  2. A. Gnagey

    Present address: Department of Biology, Case Western Reserve University, 44106, Cleveland, OH, USA

Authors and Affiliations

  1. Department of Developmental Genetics and Anatomy, School of Medicine, Case Western Reserve University, 44106, Cleveland, OH, USA

    Quiao-Yun Jiang, A. Gnagey & M. Jacobs-Lorena

  2. Department of Oral Biology, School of Dentistry, Case Western Reserve University, 44106, Cleveland, OH, USA

    B. Tandler

Authors
  1. Quiao-Yun Jiang
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  2. A. Gnagey
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  3. B. Tandler
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  4. M. Jacobs-Lorena
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Jiang, QY., Gnagey, A., Tandler, B. et al. Isolation of plasma membranes from Drosophila embryos. Mol Biol Rep 11, 19–24 (1986). https://doi.org/10.1007/BF00417590

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  • DOI: https://doi.org/10.1007/BF00417590

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