Craig S, Moore P J, Dunahay T G
Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder 80309-0347.
Scanning Microsc. 1987 Sep;1(3):1431-7.
This paper illustrates post-embedding immunogold labelling of protein and polysaccharide molecules of plant cells. For EM studies, one is restricted (for most plant cells) to the post-embedding approach because the surrounding cell wall prevents access of antibodies (and secondary gold-tagged markers) to internal sites. The large size of many plant cells also does not lend itself to diffusional entry of antibodies. The molecules localized include seed storage proteins that are large and present in major quantities, a smaller less abundant, water soluble albumin, an oxygen-binding protein, components of the photosynthetic electron transport chain, and complex sugars from the cell wall. A range of preparative procedures and embedding plastics are used.
本文阐述了植物细胞蛋白质和多糖分子的包埋后免疫金标记。对于电子显微镜研究而言(对于大多数植物细胞来说),由于周围的细胞壁会阻止抗体(以及二级金标记物)进入内部位点,所以只能采用包埋后方法。许多植物细胞体积较大,这也不利于抗体通过扩散进入细胞。被定位的分子包括大量存在的大种子贮藏蛋白、一种含量较少的较小水溶性白蛋白、一种氧结合蛋白、光合电子传递链的组分以及细胞壁中的复合糖。使用了一系列制备程序和包埋塑料。