|Immunofluorescence (Frozen)||1:50 - 1:200|
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Recommended Fluorochrome-conjugated Anti-Rabbit secondary antibodies:
NOTE: When using any primary or fluorochrome-conjugated secondary antibody for the first time, titrate the antibody to determine which dilution allows for the strongest specific signal with the least background for your sample.
Cover sections with 4% formaldehyde dilute in 1X PBS.
NOTE: Formaldehyde is toxic, use only in fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised July 2016
Protocol Id: 151
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro300 of human dopamine D(2) receptor/D2R protein.
Dopamine is a neurotransmitter that plays important roles in the brain, particularly in dopamine pathways that control the motivational component of reward-motivated behavior. These behavioral outputs are generated by the basal ganglia via its interaction with multiple brain areas that modulate sensorimotor, emotional, and cognitive information (1). The brain’s major dopaminergic input is into the striatum, a region of the basal ganglia composed of GABAergic medium spiny neurons (MSNs). Two major subpopulations of MSN exist in the striatum that are distinguished by the expression of dopamine receptor subtypes, the dopamine D(1) receptor subtype and the dopamine D(2) receptor subtype (D1R and D2R, respectively) (2,3). As a family of proteins, dopamine receptors are a class of G protein-coupled receptors (GPCRs) consisting of 5 subtypes that, upon initiation, drive downstream signaling cascades that modulate neuronal function (1). Dopamine receptors form homo- and hetero-multimers with subunits within their protein family but also with other GPCRs, including Adenosine Receptor A2a, suggesting that dopamine receptor activity might be finely tuned and altered under certain conditions (4). Dopamine receptors have been studied as a therapeutic target for several neuropsychiatric and developmental disorders, as well as neurodegenerative diseases, including Parkinson’s disease (5-8). Dopamine receptors are also expressed outside of the brain and may have diverse functions beyond the central nervous system, including regulating innate and adaptive immunity (9).
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