安定細胞株製品 » Human Recombinant D1 Dopamine Receptor Stable Cell Line
CHO-K1/D1 Stable Cell Line

This cell-based assay is based on HTRF® technology (Homogeneous Time-Resolved Fluorescence). It is a competitive immunoassay that uses cAMP labeled with the d2 acceptor flourophore and an anti-cAMP monoclonal AB labeled with Europium Cryptate. The FRET signal decreases as cAMP concentration rises.
Agonist Assay Protocol
1. Seed 5 μl CHO-K1/D1 cells into a 384-well low volume plate, 3,000 cells per well.
2. Add 5 μl compound or dopamine (diluted in buffer) to each well and incubate the plate for 30 min at 23°C.
3. Add 5ul of cAMP-d2 conjugate solution to each well.
4. Add 5μl of cAMP-AB lysis buffer solution to each well.
5. Incubate the plate in the dark for one hour at 23°C.
6. Read the plate PHERAstar PLUS (BMG Labtech, Offenburg, Germany).

CHO-K1/D1 Stable Cell Line

Dopamine is the predominant catecholamine neurotransmitter found in mammalian brain, where it controls a variety of functions including locomotor activity, cognition, emotion, positive reinforcement, food intake, and endocrine regulation. It also plays multiple roles in the periphery as a modulator of cardiovascular function, catecholamine release, hormone secretion, vascular tone, renal function, and gastrointestinal motility. The dopamine receptor family consists of five members, which are classified into two groups, D1-like (D1 and D5) and D2-like (D2, D3, and D4).
M00247
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Description

Dopamine is the predominant catecholamine neurotransmitter found in mammalian brain, where it controls a variety of functions including locomotor activity, cognition, emotion, positive reinforcement, food intake, and endocrine regulation. It also plays multiple roles in the periphery as a modulator of cardiovascular function, catecholamine release, hormone secretion, vascular tone, renal function, and gastrointestinal motility. The dopamine receptor family consists of five members, which are classified into two groups, D1-like (D1 and D5) and D2-like (D2, D3, and D4).
Synonyms

D1 receptor, D1A, DADR, DRD1A, Gpcr15, D1A, Drd-1, Drd1, D(1A) dopamine receptor, Dopamine-1A receptor, dopamine D1 receptor, dopamine receptor 1A, dopamine receptor D1A, D1 receptor, C030036C15Rik, Drd1a

Overview
Applications Functional assay for D1 receptor

Product Introduction
Storage Liquid nitrogen immediately upon delivery
Species Human

Culture Conditions
Freeze Medium 45% culture medium, 45% FBS (Cat. #10099-141, Gibco), 10% DMSO (Cat. #D2650, Sigma)
Culture Medium Ham’s F-12 K (Kaighn’s), 10% FBS, 200 µg/ml Zeocin (Cat. #R250-01, Life Technologoes)

Examples
  • CHO-K1/D1 Stable Cell Line
    • CHO-K1/D1 Stable Cell Line

    This cell-based assay is based on HTRF® technology (Homogeneous Time-Resolved Fluorescence). It is a competitive immunoassay that uses cAMP labeled with the d2 acceptor flourophore and an anti-cAMP monoclonal AB labeled with Europium Cryptate. The FRET signal decreases as cAMP concentration rises.
    Agonist Assay Protocol
    1. Seed 5 μl CHO-K1/D1 cells into a 384-well low volume plate, 3,000 cells per well.
    2. Add 5 μl compound or dopamine (diluted in buffer) to each well and incubate the plate for 30 min at 23°C.
    3. Add 5ul of cAMP-d2 conjugate solution to each well.
    4. Add 5μl of cAMP-AB lysis buffer solution to each well.
    5. Incubate the plate in the dark for one hour at 23°C.
    6. Read the plate PHERAstar PLUS (BMG Labtech, Offenburg, Germany).