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Human CHRM1 Activation 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 for the activation of Human Cholinergic Receptor, Muscarinic 1 (CHRM1). INDIGO’s CHRM1 reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the CHRM1. In addition to CHRM1 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 CHRM1. 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 ModeActivation
Kit Components
  • CHRM1 Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Acetylcholine (chloride) (20 mM in DMSO)
  • Detection Substrate
  • Detection Buffer
  • 96-well, collagen-coated assay plate (white, sterile, cell-culture ready)
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Activation of CHRM1. Reporter cells were treated with the reference activators Acetylcholine (chloride) (provided), BQCA (a selective positive allosteric modulator of CHRM1), Carbamoylcholine (chloride), Pilocarpine (chloride), Acetyl-B-methylcholine (chloride), and Bethanechol (chloride) for 5 h after an overnight pre-incubation. All chemicals were procured from Cayman Chemical (Ann Arbor, MI, USA). Luminescence was quantified and values of average (n = 4) RLU, standard deviation (SD), Fold-Activation, and Z’ values were calculated. The least-squares method of non-linear regression was used to plot activity changes vs. Log10 [Compound, nM], and EC50 values were determined, using GraphPad Prism software. The absence of signal in Acetylcholine (chloride) treated ‘Mock’ cells (which express the NFAT-Luc reporter vector, but do not express CHRM1) confirms that the observed ligand-dependent response is specific to CHRM1 activation.

Target Background

Muscarinic acetylcholine receptors are members of the G-protein-coupled receptor (GPCR) superfamily that respond to the endogenous neurotransmitter acetylcholine. They are involved in a wide range of processes in the central and peripheral nervous system, which makes them of interest as drug targets for conditions including Alzheimer’s disease, schizophrenia, and drug addiction. The muscarinic acetylcholine receptor subfamily is composed of five subtypes, known as M1 – M5, and by their corresponding genes names (i.e., CHRM1 – CHRM5). However, targeting a specific subtype has proved challenging due to the high degree of homology of the orthosteric binding site for acetylcholine.

The CHRM1 subtype is expressed in the cerebral cortex and hippocampus regions of the brain, pointing to its key role in cognitive function. Targeting the less conserved allosteric binding sites of CHRM1 have resulted in the development of selective activators of this receptor enabling the specific targeting of the M1 subtype. These opportunities for novel drug development ensure that M1 remains a viable therapeutic target.

Citations

Also available as a service

Muscarinic 1 Activation (CHRM1)

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