title: Poly- and monoclonal antibodies against recombinant rat brain calbindin D-28K were produced to map its selective distribution in the central nervous system.
abstract: 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.
authors.affiliation: Department of Pediatrics, University of Zurich, Switzerland.
authors.forename: M R
authors.initials: MR
authors.lastname: Pinol
authors.forename: U
authors.initials: U
authors.lastname: Kägi
authors.forename: C W
authors.initials: CW
authors.lastname: Heizmann
authors.forename: B
authors.initials: B
authors.lastname: Vogel
authors.forename: J M
authors.initials: JM
authors.lastname: Séquier
authors.forename: W
authors.initials: W
authors.lastname: Haas
authors.forename: W
authors.initials: W
authors.lastname: Hunziker
mesh_terms: D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001921:Brain / Q000378:metabolism*; D064026:Calbindins; D002645:Chickens; D004247:DNA; D007106:Immune Sera / Q000276:immunology; D007150:Immunohistochemistry; D007668:Kidney / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D051381:Rats; D011919:Rats, Inbred Strains; D011994:Recombinant Proteins; D064030:S100 Calcium Binding Protein G / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D014018:Tissue Distribution; D014808:Vitamin D Deficiency / Q000378:metabolism
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D064026:Calbindins; D007106:Immune Sera; D011994:Recombinant Proteins; D064030:S100 Calcium Binding Protein G; D004247:DNA