Isolation and immunohistochemical localization of a cerebellar protein

Our previous report described that a protein called spot 35 is found in the cerebellar cytosol of adult rats by two-dimensional gel electrophoresis. In this paper we isolated this protein from the soluble fraction of bovine and rat cerebella and then prepared an antiserum against the bovine protein. This protein shows pI around 5.3 and Mr around 27 kdalton. Determination of the amino acid composition of this protein shows high glutamic acid, aspartic acid and leucine contents. Using the antiserum we examined the immunohistochemical localization of this protein by the peroxidase-antiperoxidase method. Purkinje cells, their dendrites and axons were immunohistochemically stained in the cerebella of adult rats, rabbits and humans. Other cells, such as granule cells and glial cells, and myelin did not react to the antiserum.