M4 Muscarinic Acetylcholine Receptor Assay Cell Line

[CAT#: NCL2110P278]

Cell Types:: Other Cells

Certificate of Analysis Lookup
To download a Certificate of Analysis, please enter a lot number in the search box below. Note: Certificate of Analysis not available for kit components.
Lot Number

Inquiry

Product Overview

Description

Cell line information: CHRM4 PKA Maxensor cell line
Official Name: Muscarinic Acetylcholine Receptor M4
DNA Accession Number: X15265
Host Cell: U2OS
Resistance: G418 + Puromycin
Quantity:> 3 x 10^6 cells / vial
Storage: Liquid Nitrogen

Features

The red PKA Maxensor-CHRM4 contains U2OS cells stably expressing the red PKA Maxensor biosensor and muscarinic acetylcholine receptor M4 (unlabeled). Creative Biolabs' red PKA Maxensor-CHRM4 cell line is designed to test compounds or analyze their ability to modulate muscarinic acetylcholine receptor M4. When the agonist binds to CHRM4, Gi/o is activated, which in turn triggers a cellular response involving PKA activation. The activated PKA interacts with the PKA Maxensor biosensor to increase its fluorescence intensity. This cell line has been verified by measuring the PKA activation in the cytosol and analyzing the intensity and intracellular distribution of the red PKA Maxensor biosensor. Using Oxotremorine as an agonist to analyze changes in fluorescence intensity and biosensor redistribution, this highly reproducible measurement has been validated.

Cell Types

Other Cells

Application Notes

About the red PKA Maxensor biosensor
The red PKA Maxensor biosensor is a fluorescent peptide. When PKA is activated, it will change its position in the cell and increase its own fluorescence intensity in a dose response manner. Before PKA is activated, the fluorescent biosensor is located in the cell membrane. The activated PKA interacts with the PKA Maxensor biosensor, resulting in changes in its structural folding and promoting its cellular relocation in cell vesicle transport. In cell lines co-expressing the red PKA Maxensor biosensor and the GPCR of interest, the activity of living cells can be easily quantified by fluorescence granularity image analysis or fluorescence intensity analysis.

Research Areas

GPCR

Assay

M4 Muscarinic Acetylcholine Receptor Assay

Assay Description

The M4 Muscarinic Acetylcholine Receptor Assay (CHRM4) from Creative Biolabs uses Maxensor Biosensors technology to measure CHRM4 receptor activation. CHRM4 signaling occurs through adenylate cyclase inhibition of the trimer Gi/o protein and is reduced through PKA activation cAMP.

In this assay, we use Maxensor Biosensors technology to measure receptor activation/inhibition through PKA activation. The high reproducibility of this assay allows monitoring the activation of M4 muscarinic acetylcholine receptors in living cells.

Assay Characteristics

Readout: cAMP Flux (Gi/o activation)

Agonist: Oxotremorine

EC50 Agonist: 4.72 x 10⁶-8 M for Gi/Go Activation

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.