"RXFP4" Related Products

Human RXFP4 Stable Cell Line-CHO Galpha16

Cat.No.: CSC-RG0930
Cell Line Description: CHO Galpha16-HuRXFP4 cell line is clonally-derived from a CHO cell line, which has been transfected with a human relaxin/insulin-like family peptide receptor 4 RXFP4 to allow expression of the human RXFP4. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: The RXFP4 receptor belongs to the Relaxin Family Peptide Receptors family. It was originally cloned as orphan receptor GPR100 and GPR142 but subsequently has been shown to be the physiological target for INSL3 (insulin-like peptide 3); it also responds to relaxin-3 at high affinity. Human RXFP4 is expressed in heart, skeletal muscle, kidney, liver and placenta and has been shown to be Gi-coupled. This cell line couples via Ga16 to the calcium pathway. Although to our knowledge RXFP4 has yet to be used as a drug discovery target, this cell line will provide utility for counter-screening compounds targeted to other members of this family that may have therapeutic utility for a wide range of diseases including cardiac and renal failure (RXFP1), asthma (RXFP1), scleroderma (RXFP1), infertility and contraception (RXFP2), anti-anxiety and anti-obesity (RXFP3).
Growth Properties: Adherent
Morphology: Epithelial-like
Host Cell: CHO-K1
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific expression of human RXFP4.2. Protein expression: RXFP4 expression in this cell line has been validated by WB.3. Functional validation: binding, calcium assay.
Sub-type: Relaxin
Propagation: Complete growth medium: Ham"s F12, 10% FBS, 1% NEAA, 400 μg/mL G418, 250 μg/mL Hygromycin BAtmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Starting Cells From Frozen Cell Stock: 1. Remove the packaging cell lines from liquid nitrogen and carry out a quick thaw. Float the cells in the 37°C water bath for 2 minutes until nearly (80%) thawed. Once cells are thawed, it is important to dilute the cells 1:10 in growth media immediately to reduce the potentially toxic effects of the DMSO preservative on the cells.2. Clean the outside of the vial with 70% ethanol before opening.3. Remove the cells from the vial and add slowly into a 15ml conical tube containing 10 ml pre-warmed media. 4. Centrifuge for 3 minutes 1000 xg to pellet cells and remove the supernatant.5. Add 14 ml of media and transfer cells to a T25 flask or a 100 mm culture dish.6. Place the cells in the 37°C incubator with 5% CO2.7. Allow incubation for 3-4 days to reach confluence. The cells will re-attach to the surface over a period of several days in culture at 37°C.
Subculturing: 1. Remove and discard culture medium.2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.5. Add appropriate aliquots of the cell suspension to new culture vessels.Incubate cultures at 37°C.Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended Medium Renewal: 2 to 3 times per week
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: RXFP4 relaxin/insulin-like family peptide receptor 4 [ Homo sapiens ]
Official Symbol: RXFP4
Synonyms: RXFP4; relaxin/insulin-like family peptide receptor 4; G protein coupled receptor 100 , GPR100, relaxin 3 receptor 2 , relaxin family peptide receptor 4 , RLN3R2; relaxin-3 receptor 2; GPCR142; RXFPR4; RLN3 receptor 2; relaxin 3 receptor 2; G protein-coupled receptor 100; G-protein coupled receptor 100; relaxin family peptide receptor 4; G-protein coupled receptor GPCR142; insulin-like peptide INSL5 receptor; GPR100; RLN3R2; MGC126556; MGC126558;
Gene ID: 339403
mRNA Refseq: NM_181885
Protein Refseq: NP_871001
MIM: 609043
UniProt ID: Q8TDU9
Chromosome Location: 1q22
Pathway: Class A/1 (Rhodopsin-like receptors), organism-specific biosystem; G alpha (i) signalling events, organism-specific biosystem; GPCR downstream signaling, organism-specific biosystem; GPCR ligand binding, organism-specific biosystem; Peptide ligand-binding receptors, organism-specific biosystem; Relaxin receptors, organism-specific biosystem; Signal Transduction, organism-specific biosystem;
Function: G-protein coupled receptor activity; angiotensin type II receptor activity; receptor activity; signal transducer activity;
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