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).

產品編號	M00247
規(guī)格	2 vials 大單詢價
數(shù)量
價格	詢價


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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

產品簡介

Storage

Liquid nitrogen immediately upon delivery

培養(yǎng)條件

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

圖例

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).

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CHO-K1/D1 Stable Cell Line

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