"CRHR2" Related Products


Human CRHR2 Stable Cell Line-HEK293

Cat.No.: CSC-RG0009
Cell Line Description: HEK293-HuCRHR2 cell line is a hypotriploid human cell line, which has been transfected with a Human Corticotropin Releasing Hormone Receptor 2(CRHR2) to allow stably express of the human CRHR2. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: Corticotropin Releasing Hormone Repceptor 2 is the gene that encodes a protein also known as CRHR-2. The Corticotropin Releasing Hormone Receptor family is a group of Gcoupled receptors that shows high affinity for Corticotropin Releasing Hormone (CRH). CRH is a polypeptide hormone and neurotransmitter synthesized in the hypothalamus that is the principal neuroregulator of the hypothalamic-pituitaryadrenocortical axis. CRH is involved in the stress response and the stimulation of the pituitary synthesis of ACTH (Adrenocorticotropic hormone).
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: HEK293
Cell Line Validation: 1. Gene expression: RT-PCR experiments determined specific expression of human CRHR2.2. Functional validation: Corticotropin Releasing Factor dose response in calcium assay; CRF-2 dose response curve in cAMP assay.
Sub-type: Corticotropin-Releasing Factor
Propagation: Complete growth medium: EMEM (EBSS), 2mM Glutamine, 1% Non Essential Amino Acids (NEAA), 10% FCSAtmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Starting Cells From Frozen Cell Stock: 1. Prepare a poly-L-lysine coated flask (2μg/cm2, T-75 flask is recommended). Add 10 ml of sterile water to a T-75 flask and then add 15 μl of poly-Llysine stock solution (10 mg/ml). Leave the flask in incubator overnight (minimum one hour at 37°C incubator). 2. Prepare complete medium: decontaminate the external surfaces of medium and medium supplements with 70% ethanol and transfer themto sterile field. Aseptically open each supplement tube and add them to the basal medium with a pipette. Rinse each tube with medium to recover the entire volume.3. Rinse the poly-L-lysine coated flask with sterile water twice and add 20 ml of complete medium to the flask. Leave the flask in the hood and go to thaw the cells.4. Place the vial in a 37°C waterbath, hold and rotate the vial gently until the contents are completely thawed. Remove the vial from the waterbath immediately, wipe it dry, rinse the vial with 70% ethanol and transfer it to a sterile field. Centrifuge at 125xg for 5 min to collect the cells Remove the cap, being careful not to touch the interior threads with fingers. Remove the frozeen media using 1 ml eppendorf pipette and gently resuspend the contents of the vial in 1ml of pre-warmed complete growth media.5. Dispense the contents of the vial into the equilibrated, poly-L-lysine coated culture vessels containing 9ml prewarmed complete growth media. A seeding density of 3 x 10^6 cells/cm2 is recommended. 6. Place the culture vessels to the incubator.7. For best result, do not disturb the culture for at least 16 hours after the culture has been initiated. Change the growth medium the next day to remove the residual DMSO and unattached cells, then every other day thereafter. A healthy culture will display stellate or spindle-shaped cell morphology, nongranular cytoplasm, and the cell number will be double after two to three days in culture.
Subculturing: 1. Subculture the cells when they are over 90% confluent.2. Prepare poly-L-lysine coated cell culture flasks.3. Warm medium, trypsin/EDTA solution, trypsin neutralization solution, and DPBS to room temperature. We do not recommend warming the reagents and medium at 37°C waterbath prior to use.4. Rinse the cells with DPBS.5. Incubate cells with 5 ml of trypsin/EDTA solution (in the case of T-75 flask) until 80% of cells are rounded up (monitored with microscope). Add 5ml of trypsin neutralization solution to the digestion immediately and gently rock the culture vessel. 6. Harvest and transfer released cells into a 50 ml centrifuge tube. Rinse the flask with another 10 ml of growth medium to collect the residue cells. Examine the flask under microscope to make sure the harvesting is successful by looking at the number of cells left behind. There should be less than 5%. 7. Centrifuge the harvested cell suspension at 1000 rpm for 5 min and resuspend cells in growth medium.8. Count cells and plate them in a new, poly-L-lysine coated flask with cell density as recommended.
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Complete growth medium 90%; DMSO, 10%
Storage: Liquid nitrogen
Preservation: 1. Detach cells from culture dish according to the Sub-Culture Procedure.2. Resuspend cells at a density of 5 x 10^6 cells/mL in freeze medium.Note: A T-75 culture flask typically yields enough cells for preparing two frozen vials.3. Aliquot 1 mL cells into cryogenic vials.4. Place vials in a freezing container and store at –80 °C overnight.5. Transfer vials to liquid nitrogen for long term storage. If properly stored, cells should remain stable for years.
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: CRHR2 corticotropin releasing hormone receptor 2 [ Homo sapiens ]
Official Symbol: CRHR2
Synonyms: CRHR2; corticotropin releasing hormone receptor 2; corticotropin-releasing factor receptor 2; CRF RB; CRF2; HM CRF; CRH-R2; CRF2 receptor, beta isoform; CRFR2; CRF-RB; HM-CRF;
Gene ID: 1395
mRNA Refseq: NM_001202475
Protein Refseq: NP_001189404
MIM: 602034
UniProt ID: Q13324
Chromosome Location: 7p21-p15
Pathway: Class B/2 (Secretin family 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 B Secretin-like, organism-specific biosystem; Neuroactive ligand-receptor interaction, organism-specific biosystem; Neuroactive ligand-receptor interaction, conserved biosystem;
Function: corticotrophin-releasing factor receptor activity; protein binding; receptor activity; signal transducer activity;

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