cAMP ELISA
The level of cAMP in the hippocampus was measured by a direct
competitive ELISA kit (Abcam, USA) as directed by the manufacturer.
After weighing the frozen tissue, 0.1M HCl was added in a 5X volume.
Then the sample was homogenized on ice and centrifuged at high speed for
five minutes, collecting the supernatant. After loading the samples into
96-well plates, the samples were incubated for an hour with the cAMP
antibody. The wells were incubated for an additional hour with cAMP-
Horseradish Peroxidase (HRP) and washed three times with wash buffer
before the final step. Thereafter, HRP developer was added and incubated
for 1 hour. A 1 M HCl solution was used to stop the reaction, and the
color developed was measured using an ELx800 plate reader (BioTek, USA).