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Poly‐ and Monoclonal Antibodies Against Recombinant Rat Brain Calbindin D‐28K Were Produced to Map Its Selective Distribution in the Central Nervous System
Author(s) -
Pinol M. Rosa,
Kägi Urs,
Heizmann Claus W.,
Vogel Brigitte,
Séquier JeanMarc,
Haas Werner,
Hunziker Willi
Publication year - 1990
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1990.tb04879.x
Subject(s) - calbindin , polyclonal antibodies , antiserum , calretinin , microbiology and biotechnology , western blot , biology , recombinant dna , monoclonal antibody , immunohistochemistry , blot , antibody , immunocytochemistry , calcium binding protein , calcium , chemistry , biochemistry , endocrinology , immunology , gene , organic chemistry
Many processes in the CNS depend on calcium. The calcium signal is transduced into an intracellular response via Ca 2+ ‐binding proteins, including calbindin D‐28K. In many laboratories, polyclonal antibodies against chicken in testinal 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 D28K 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.