The production of antibodies that distinguish rat choline acetyltransferase from its splice variant product of a peripheral type.
To produce antibodies that permit the immunohistochemical discrimination of choline acetyltransferase of the common type (cChAT) from its splice variant of a peripheral type (pChAT), we immunized rabbits with a cChAT specific recombinant protein encoded by ChAT exons 7 and 8 of the rat cChAT gene. Successful antibody production was proved by Western blotting on rat brain and on HEK293 cells expressing green fluorescent protein (GFP), cChAT-GFP and pChAT-GFP. By immunohistochemistry our antiserum clearly labeled known cholinergic structures in rat brain, but gave no positive staining in the trigeminal ganglion which contained many neurons positive with pChAT antiserum.
Kimura
Shin
S
Molecular Neuroscience Research Center, Shiga University of Medical Science, Otsu 520-21, Japan.
Bellier
Jean-Pierre
JP
Matsuo
Akinori
A
Tooyama
Ikuo
I
Kimura
Hiroshi
H
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000917:Antibody Formation; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D002795:Choline O-Acetyltransferase / Q000276:immunology*; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D051381:Rats
D000906:Antibodies; D017931:DNA Primers; D002795:Choline O-Acetyltransferase