作者
Diane L Rosin, Anna Robeva, Robin L Woodard, Patrice G Guyenet, Joel Linden
发表日期
1998/11/16
期刊
Journal of Comparative Neurology
卷号
401
期号
2
页码范围
163-186
出版商
John Wiley & Sons, Inc.
简介
The A2A adenosine receptor (A2A‐AR) transcript and radioligand binding sites have a distinct distribution in rat brain, restricted primarily to the striatum, nucleus accumbens and olfactory tubercles. We describe here the use of purified recombinant human A2A‐ARs to generate a monoclonal antibody that has been used to better resolve the distribution of A2A‐ARs in rat brain. The antibody can detect 1 ng of purified recombinant receptor by Western blotting and is potent (EC50 = 0.62 μg/ ml) and highly selective for the A2A‐AR subtype. By Western blotting, the apparent molecular mass of recombinant and rat striatal receptors shifts upon deglycosylation from 43–48 to 42 kilodaltons. Analyses of chimeric A1/ A2A‐ARs and synthesis of a blocking peptide pinpointed the epitope (SQPLPGER) of the antibody to the center of the third intracellular loop of the receptor. Incubation of rat striatal membranes with antibody …
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