Alkaline phosphatase (ALP) was purified from the rat kidney. The appro
priate enzyme protein was injected subcutaneously into a rabbit as an
antigen. The obtained antiserum showed a single precipitin line in the
double diffusion test of Ouchterlony. The localization of ALP in the
rat liver was examined by means of immunohistochemical techniques. At
the light microscopic level, positive reactions were recognized along
cell borders between adjacent hepatocytes. On examination with an elec
tron microscope, gold particles were observed only on the surface of m
icrovilli of bile canaliculi by immunogold staining. On the other hand
, by the peroxidase-labeled antibody method, reaction products appeare
d not only on the bile canalicular membrane but also on the sinusoidal
and lateral membranes.