Enzyme-linked Immunosorbent Assay (ELISA)
Brains were flash frozen and stored in a -80 freezer. The PFC was removed was kept frozen throughout the entire process. The PFC was identified using a mouse brain atlas and visual landmarks. The tissue removed was within AP 2.93mm and 1.33mm. To prepare samples for ELISA, 1x phosphate-buffered saline (PBS) was micro-pipetted into the vials at a concentration of 10% tissue and 90% PBS as per recommendations of the kits. Samples were centrifuged at 1000 x g for 20 minutes and the supernatant was removed. Instructions were followed for each kit thoroughly. Each ELISA kit was purchased from MyBioSource, and this study tested for CB1R (Catalogue: MBS2533482) and 5-HT1AR (Catalogue: MBS287600).