Cat.No. : CSC-RG0069 Host Cell: CHO-K1
| Cat. No. | CSC-RG0069 |
| Background | The muscarinic cholinergic receptors belong to a larger family of G protein-coupled receptors. The functional diversity of these receptors is defined by the binding of acetylcholine and includes cellular responses such as adenylate cyclase inhibition, phosphoinositide degeneration, and potassium channel mediation. Muscarinic receptors influence many effects of acetylcholine in the central and peripheral nervous system. The clinical implications of this receptor are unknown; however, stimulation of this receptor is known to increase cyclic AMP levels. |
| Gene | CHRM5 |
| Gene Species | Homo sapiens (Human) |
| Alias | CHRM5, HM5, MGC41838 |
| Host Cell | CHO-K1 |
| Species | Cricetulus griseus (Chinese hamster) |
| Morphology | Epithelial-like |
| Stability | Validated for at least 10 passages |
| Application | 1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Research on the mechanisms of GPCR-related diseases |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Growth Properties | Adherent |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
A: The Human CHRM5 Stable Cell Line-CHO (Chinese Hamster Ovary cell line stably expressing human muscarinic acetylcholine receptor M5) plays a pivotal role in drug screening, especially in the development of drugs targeting the muscarinic acetylcholine receptors. This cell line enables researchers to simulate the interaction between the receptor and drug molecules in vitro, allowing for the screening of potential agonists or antagonists. This is significant for the treatment of neurodegenerative diseases related to the cholinergic system, such as Alzheimer's and Parkinson's diseases.
A: Applications of Human CHRM5 Stable Cell Line-CHO in studying muscarinic receptor signaling include but are not limited to: 1) analyzing the pharmacological properties of the CHRM5 receptor, such as affinity, the effects of agonists and antagonists; 2) investigating the downstream signaling pathways activated by the receptor, such as G protein coupling and second messenger systems; 3) exploring the conformational changes and drug binding mechanisms of the receptor; 4) assessing functional changes of the receptor in disease states. These studies contribute to a deeper understanding of the physiological and pathological processes of the cholinergic system.
A: To utilize Human CHRM5 Stable Cell Line-CHO for high-throughput screening (HTS), an assay based on the activity of the CHRM5 receptor, such as fluorescence resonance energy transfer (FRET) or calcium imaging, needs to be established. Then, the cell line is seeded into microplate wells, with a small number of cells per well. Automated addition of a series of drug candidates to each well is followed by monitoring changes in receptor activity. By analyzing this data, potential drug candidates can be rapidly identified. This method significantly increases screening efficiency and accelerates the drug discovery process.
A: To optimize the experimental design for studying the ligand selectivity of muscarinic receptors using Human CHRM5 Stable Cell Line-CHO, several key steps should be included: First, ensure that the cell line is cultured under optimal growth conditions to maintain the stability and activity of the receptor. Second, select an appropriate assay method, such as radioligand binding assays or functional assays, to evaluate the affinity and efficacy of ligands with the receptor. Additionally, design experiments with different concentrations of ligands to determine their saturation and maximal response with the receptor. Finally, perform statistical analysis, such as Scatchard analysis or Hill plot, to quantitatively analyze the interaction between ligands and receptors. These steps help to accurately measure ligand selectivity, providing important information for drug design.
A: When studying the structure-activity relationship (SAR) of muscarinic receptors using Human CHRM5 Stable Cell Line-CHO, a series of drug molecules with structural variations are assessed. Initially, a range of drug molecules with systematic structural changes are selected. Then, functional assays, such as intracellular calcium release or cAMP production, are performed using the cell line to evaluate the activity of these molecules on the CHRM5 receptor. By comparing the activity of drug molecules with different structures, the impact of structural changes on receptor activity can be revealed, thus establishing a structure-activity relationship. This approach helps to understand how drug molecules interact with the receptor and guides the design of more effective drugs.
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