M5 Muscarinic Acetylcholine Receptor Assay Cell Line

[CAT#: NCL2110P277]

Cell Types:: Other Cells

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Product Overview

Description

Cell line information: DAG Maxensor M5 Cell Line
DNA Accession Number: Gene Bank M80333
Official Full Name: Muscarinic acetylcholine receptor M5
Host Cell: DAG Maxensor U2OS Cell Line
Fluorescence: excitation/emission maxima at 574/602 nm
Resistances: Puromycin & G418
Quantity:> 3 x 10^6 cells / vial
Storage: Liquid Nitrogen

Features

The DAG Maxensor M5 cell line contains 3 million U2OS cells, which stably express DAG Maxensor biosensor and human M5 receptor. The Creative Biolabs DAG Maxensor M5 cell line is designed to determine compounds or analyze their ability to modulate M5 receptors. When the agonist binds to M5, the G protein is activated, which in turn triggers a cellular response mediated by the second messenger (DAG). This cell line has been verified by measuring the increase in DAG in the cytosol and analyzing the increase in fluorescence of the DAG Maxensor biosensor in the cell. This cell line allows image analysis of compound-induced stimuli. The use of Oxotremorine as an M5 agonist in high-content analysis (HCA) and fluorescence plate readers validated this highly reproducible assay.

Cell Types

Other Cells

Application Notes

About Maxensor DAG Maxensor Biosensor:
DAG Maxensor biosensor is a red fluorescent peptide that increases the fluorescence intensity in cells in the presence or absence of DAG. The increase in the concentration of the second messenger leads to changes in the folding of the Maxensor biosensor structure, thereby promoting its cell fluorescence intensity. In cell lines co-expressing DAG Maxensor biosensor and GPCR, after activating the receptor with its agonist, the activity of living cells can be easily quantified by fluorescence increase signal. The DAG Maxensor biosensor has true red fluorescence (excitation/emission maximum values of 574/602 nm respectively), which is most suitable for detection through the most popular filter sets and can be easily distinguished from background signals.

Research Areas

GPCR

Assay

M5 Muscarinic Acetylcholine Receptor Assay

Assay Description

The M5 Muscarinic Acetylcholine Receptor Assay (CHRM5) from Creative Biolabs allows compounds to be tested or analyzed for their ability to modulate CHRM5. When the ligand binds to CHRM5, it activates the G protein; this in turn triggers calcium Mediated cell response. The combination of agonist and CHRM5 induces intracellular calcium mobilization and PKC activation through PLC-IP3 and DAG pathways, respectively.

The M5 muscarinic acetylcholine receptor test involves the measurement of diacylglycerol (DAG) in the cytosol through PKC activation. The high reproducibility of this assay allows monitoring the activation of CHRM5 receptors in living cells.

Assay Characteristics

Readout: Diacyl Flux

Agonist: Oxotremorine

EC50 Agonist: 1.48 x 10⁶-8 M for DAG Flux

Type of Assay: Cell-Based/Functional

Detection Method: Fluorimetry

Properties

Form

Frozen

Cell Purity

>95%

Cell Viability

>90%

Mycoplasma Testing

The cell line has been screened using the luciferase based mycoplasma detection kit to confirm the absence of mycoplasma species.

Sterility Testing

Sterility testing was performed in accordance with USP and EP regulations. All of our sterility testing is performed in an isolator or clean room environments. The cell line has been screened using the membrane filtration testing methods to confirm the absence of aerobic, anaerobic and fungi microorganisms.

Genetic Stability Testing

Cell genetic stability study was perfomed under ICH guidelines. We provide guidance on the appropriate testing program upon your requirements.

Shipping

Dry ice

Storage

Liquid Nitrogen

Handling Advice

Frozen cells:Upon receipt, frozen ampoules should be transferred directly to gaseous phase liquid nitrogen without delay, unless they are to be used straight away.

Research Use Only

For research use only, not for diagnostic or therapeutic use.

Warnings

Store under recommended storage conditions (liquid nitrogen). Do not expose to high temperature. After expiration, discard all remaining reagents. It is recommended to use cells within ten generations.

Quality Control

No bacteria, yeast, fungi, mycoplasma, virus

Target Details

Target

M5/CHRM5

Official Name

CHRM5

Full Name

M5 Muscarinic Acetylcholine Receptor

Alternative Names

M5 receptor; m5; acetylcholine receptor; muscarinic 5; cholinergic receptor; muscarinic 5; muscarinic acetylcholine receptor M5; muscarinic acetylcholine receptor 5; M5R

Publications