Dopamine transporter (DAT) plays the most important role in terminating the
actions of dopamines released into the synaptic cleft. DAT is also the tar
get of various psychotropic drugs such as cocaine and amphetamine. In this
study we prepared DAT-specific antibodies using the 2nd extracellular loop
of rat BAT as an antigen. The 2nd extracellular loop of the rat DAT was exp
ressed in bacteria as a fusion protein with glutathione-S-transferase, and
injected into rabbits to raise antibodies. Produced antibodies clearly reco
gnized the rat DAT in ELISA, immunoblotting, and immunoprecipitation. As ex
pected from the high sequence homology between the rat and human DAT, the a
ntibodies raised for the rat DAT crossreacted with the human DAT in the imm
unoblotting. Considering the specificity for DAT with wide range of applica
tions such as ELISA, immunoblotting, and immunoprecipitation, these antibod
ies would be valuable tool for understanding the pharmacological actions of
dopamine transporter and drug addiction.