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Human DRD1 Reporter Assay Kit

SIZE SKU PRICE
1 x-96 well format assays
3 x-32 assays in-96 well format
SIZE SKU
3 x-32 assays in-96 well format
1 x-96 well format assays

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the the Human Dopamine Receptor D1 (DRD1). INDIGO’s DRD1 reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the DRD1. In addition to DRD1 Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist, Luciferase Detection Reagent, and a cell culture-ready assay plate. The principal application of this assay is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against DRD1. This kit provides researchers with clear, reproducible results, exceptional cell viability post-thaw, and consistent results lot to lot. Kits must be stored at -80C. Do not store in liquid nitrogen. Note: reporter cells cannot be refrozen or maintained in extended culture.

Features

  • Clear, Reproducible Results

  • All-Inclusive Assay Systems
  • Exceptional Cell Viability Post-Thaw
  • Consistent Results Lot to Lot

Product Specifications

Target TypeGPCR
SpeciesHuman
Receptor FormHybrid
Assay ModeAgonist, Antagonist
Kit Components
  • DRD1 Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Dopamine HCl (40 mM)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Activation of DRD1. Activation assays were performed using the reference activator Dopamine HCl (provided), A-77636 HCl, Rotigotine and (S)-Pramipexole HCl (all from Cayman Chemical, Ann Arbor, MI).
Inhibition of DRD1. DRD1 reporter cells were co-treated with an EC80 concentration of the reference activator Dopamine HCl and varying concentrations of DRD1 inhibitors Fluphenazine HCl, SCH 39166 HBr and SKF 83566 HBr (all compounds obtained from Cayman Chemical). INDIGO’s Live Cell Multiplex (LCM) Assay confirmed that no treatment concentrations were cytotoxic (data not shown). For both Activation and Inhibition assays, luminescence was quantified and values of average (n=3) relative light units (RLU), corresponding standard deviation (SD), Fold-Activation, and Z’ values were calculated. GraphPad Prism software was used to plot data using the least-squares method of non-linear regression for Fold-Activation or RLU vs. Log10 [Cmpd, nM], and to determine EC50 / IC50 values.

Target Background

This assay utilizes proprietary human cells that provide constitutive expression of the Human Dopamine Receptor D1 (DRD1).

Dopamine acts as an essential neurotransmitter whose signaling is conducted through five G protein-coupled receptors (GPCRs), dopamine D1 to D5 receptors (DRD1-DRD5). DRD1 primarily couples to the Gαs family of G proteins to activate adenylyl cyclase and induce cAMP production.

Dysregulation of DRD1 signaling has been directly linked to many neurological diseases, for example: Parkinson’s Disease (PD), Schizophrenia, drug abuse, autism spectrum disorder (ASD) and attention deficit / hyperactivity disorder (ADHD).

PD is considered a progressive neurodegenerative disorder with increasing worldwide prevalence, and its incidence steadily increases with age. Indeed, the global burden of this disease is estimated to be more than 12 million by 2040. Dopamine agonists have demonstrated efficacy for the treatment of PD but are limited by their adverse effects. However, selective activation of DRD1 and DRD5 have been explored in small, early-phase clinical trials with some success. Although, challenges exist with pharmacokinetics and bioavailability of these test compounds. Due to their involvement in many neurological ailments, DRD1 and related receptors continue to command considerable interest in therapeutics development and drug safety screening.

Citations

Also available as a service

Dopamine Receptor D1 (DRD1)

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