Pinol M R, Kägi U, Heizmann C W, Vogel B, Séquier J M, Haas W, Hunziker W
Department of Pediatrics, University of Zurich, Switzerland.
J Neurochem. 1990 Jun;54(6):1827-33. doi: 10.1111/j.1471-4159.1990.tb04879.x.
Many processes in the CNS depend on calcium. The calcium signal is transduced into an intracellular response via Ca2(+)-binding proteins, including calbindin D-28K. In many laboratories, polyclonal antibodies against chicken intestinal calbindin D-28K have been used to study its localization in the brain (normal and degenerated) of various species, including humans, but some of these antisera cross-reacted with other proteins, including calretinin. We purified recombinant rat brain calbindin D-28K to raise antisera in rabbits and purified a recombinant rat-chicken calbindin D-28K hybrid protein to immunize mice for the generation of monoclonal antibodies. These antisera were highly specific for calbindin D-28K, as demonstrated by two-dimensional Western blotting analysis. Immunohistochemical analyses combined with in situ hybridization studies demonstrated that calbindin D-28K in the Purkinje cells of the cerebellum is independent of vitamin D. The antibodies described here will be important tools for studying the regulation of expression of calbindin D-28K and its biological function in the brain and in the PNS.
中枢神经系统（CNS）中的许多过程都依赖于钙。钙信号通过Ca2(+)结合蛋白（包括钙结合蛋白D-28K）转导为细胞内反应。在许多实验室中，针对鸡肠钙结合蛋白D-28K的多克隆抗体已被用于研究其在包括人类在内的各种物种的大脑（正常和退化的）中的定位，但其中一些抗血清与其他蛋白质（包括钙视网膜蛋白）发生交叉反应。我们纯化了重组大鼠脑钙结合蛋白D-28K以在兔中产生抗血清，并纯化了重组大鼠-鸡钙结合蛋白D-28K杂交蛋白以免疫小鼠以产生单克隆抗体。二维蛋白质印迹分析表明，这些抗血清对钙结合蛋白D-28K具有高度特异性。免疫组织化学分析与原位杂交研究相结合表明，小脑浦肯野细胞中的钙结合蛋白D-28K与维生素D无关。本文所述的抗体将成为研究钙结合蛋白D-28K在大脑和外周神经系统（PNS）中的表达调控及其生物学功能的重要工具。
