"MC4R" Related Products

Expand
Expand

Human MC4R/Galpha15 Stable Cell Line-Chem-3

Cat.No.: CSC-RG0697
Cell Line Description: This human MC4-expressing cell line is made in the Chem-3 host, which supports high levels of recombinant MC4 expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between MC4 and its ligands.
Background: The melanocortins, alpha-, beta- and -melanocyte-stimulating hormones (MSHs), are peptides derived from a precursor protein POMC. The MSH peptides bind to a family of five Gs-coupled seven transmembrane recptors (MC1-5) and play important roles in energy balance, reproductive function, pigmentation and inflammation. The activity of the melanocortins is modulated by endogenous antagonist proteins, agouti and agouti-related protein (AGRP). MC4 is expressed primarily in the CNS and appears to play a prominent role in energy homeostasis. The hypothalamus, which is a major central site for control of feeding behavior, prominently expresses MC4. Targeted deletion of MC4 in mice and naturally occurring mutations in MC4 in humans result in obese phenotypes. In addition, blockade of MC4 function by antagonists and targeted deletion of the gene in mice reverses melanocortin agonist-induced inhibition of food intake and promotes weight gain in uremia and cancer-induced cachexia.
Growth Properties: Adherent
Host Cell: Chem-3
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific silencing of MC4R. 2. Protein expression: MC4R in this cell line has been validated by immunocytochemical staining.3. EC50 for calcium mobilization by NDP-MSH: ~ 1μM.
Application: Calcium flux assay, ligand binding assays
Sub-type: Melanocortin
Propagation: Complete growth medium: RPMI-1640 containing 0.3 g/L L-glutamine, no pyruvate + 10% heat-inactivated FBS + 100 U/ml penicillin + 100 U/ml streptomycin + 1% Chem-3 Growth Supplement + G-418/Geneticin (600ug/mL)Note: The Chem-3 Growth Supplement is required for growth of Chem-3 cell linesPlating medium: RPMI-1640 containing 0.3 g/L L-glutamine, no pyruvate + 10% heat-inactivated FBS + 100 U/ml penicillin + 100 U/ml streptomycinAtmosphere: air, 95%; carbon dioxide (CO2), 5%. Temperature: 37 °C
Starting Cells From Frozen Cell Stock: 1. Immediately upon receipt, thaw cells or place cells in liquid nitrogen. Maintain frozen in liquid nitrogen for up to 5 years. 2. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37°C water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a conical tube containing 10 mL Growth Media. Centrifuge at 200 x g for 5 min. Remove the supernatant and resuspend the cell pellet in 20 mL Growth Media. Transfer to a T75 flask and place the flask in a humidified incubator at 37 degree °C with 5% CO2.
Subculturing: 1. Maintain the cells in suspension between 0.1 x 10^6 and 1 x 10^6 cells/mL. 2. Cells are typically passaged 1:10 every 2-3 days, or 1:20 every 3-4 days. Passaging ratio may be varied according to requirements of the investigator. Chem-3 cells tend to proliferate rapidly, so care should be taken not to allow them to become overconfluent.
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Freezing medium: RPMI-1640 containing 0.3 g/L L-glutamine, no pyruvate + 20% heat-inactivated FBS + 10% DMSO
Storage: Liquid nitrogen
Preservation: 1. Frozen stocks of cells should be prepared at the earliest passage possible after thawing, as follows: Count cells. Centrifuge cells at 200 x g for 5 min. Resuspend cells at 2-5 x 10^6 cells/mL in Freezing Media. Dispense 1 mL aliquots into cryopreservation vials. Freeze the cells by a controlled rate process, such as in an isopropanol-jacketed container placed at –70°C overnight. Store the vials in liquid nitrogen. 2. Cells should be passaged at least once after thawing prior to use in calcium flux assays.
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
Gene Name: MC4R melanocortin 4 receptor [ Homo sapiens ]
Official Symbol: MC4R
Synonyms: MC4R; melanocortin 4 receptor; melanocortin receptor 4; MC4-R; MGC126851; MGC138197;
Gene ID: 4160
mRNA Refseq: NM_005912
Protein Refseq: NP_005903
MIM: 155541
UniProt ID: P32245
Chromosome Location: 18q22
Pathway: Class A/1 (Rhodopsin-like receptors), organism-specific biosystem; G alpha (s) signalling events, organism-specific biosystem; GPCR downstream signaling, organism-specific biosystem; GPCR ligand binding, organism-specific biosystem; GPCRs, Class A Rhodopsin-like, organism-specific biosystem; Neuroactive ligand-receptor interaction, organism-specific biosystem; Neuroactive ligand-receptor interaction, conserved biosystem;
Function: G-protein coupled receptor activity; melanocortin receptor activity; melanocyte-stimulating hormone receptor activity; neuropeptide binding; peptide hormone binding; protein binding; receptor activity; signal transducer activity; ubiquitin protein ligase binding;

Online Inquiry

Note: There will be extra charge for optional service!

Please input "biomart" as verification code. Please review Creative BioMart's privacy policy for more information

Optional requirements on this protein    +Expand

Price Inquiry

Welcome! For price inquiries, please feel free to contact us through the form below. We will get back to you as soon as possible.