"RXFP4" Related Products

Human RXFP4/Gi/Go Stable Cell Line-CHO

Cat.No.: CSC-RG0554
Cell Line Description: These GPCR stable cell lines are non-force coupled cell lines that are validated for functional cAMP response and are fully characterized for pharmacology & specificity. Each clonal cell line is engineered to express the full length GPCR protein. These GPCR stable cell lines are used with cAMP detection kit for measuring the activation of the target (GPCR).
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 silencing of human RXFP4.2. Protein expression: RXFP4 in this cell line has been validated by immunocytochemical staining.
Application: cAMP assays
Sub-type: Relaxin Family peptide
Propagation: Complete growth medium: F12 Nutrient Mixure (HAM) + 10% FBS + 1× penicillin + 1× streptomycin + 1× L-glutamine + 800μg/ml geneticin. Atmosphere: air, 95%; carbon dioxide (CO2), 5%. Temperature: 37 °C
Starting Cells From Frozen Cell Stock: 1. Thaw frozen cells very briefly in a 37 °C water bath under sterile conditions until just before ice completely melts (30 seconds to 1 minute). Caution: Longer incubation may result in cell death.2. Remove DMSO from the media by carefully transferring thawed cells to a sterile 15 mL tube, filling tube with complete media without antibiotics pre-warmed to 37 °C, and centrifuge at 300g for 4 minutes to pellet cells.3. Resuspend cell pellet in 5 mL of pre-warmed complete media without antibiotics, transfer to a T25 flask, and grow for 24 hours. Cell recovery is greatly improved when antibiotics are omitted for the first 24 hours.4. After 24 hours, exchange with 5 mL of media containing antibiotics. Antibiotic selection must be applied after the first 24 hours or the expression of GPCR could be lost. 5. Once the cells become >70% confluent in the T25 flask, trypsinize (using a 0.05% trypsin solution) and resuspend with 5 mL of complete media. Transfer the entire cell suspension to a T75 flask containing 5 mL ofcomplete media (containing antibiotics) for continued growth.
Subculturing: 1. Remove and discard culture medium.2. Wash cells with PBS (pH=7.4) to remove all traces of serum that contains trypsin inhibitor.3. Add 2.0 ml of 0.05% (w/v) Trypsin-EDTA (GIBCO, Cat No. 25300) solution to 10 cm dish and observe the cells under an inverted microscope until cell layer is dispersed (usually within 3 to 5 minutes). Note: To avoid clumping, do not agitate the cells by hitting or shaking the dish 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, centrifuge the cells at 200g for 5min, and discard the medium.5. Resuspend the cells in culture medium and add appropriate aliquots of the cell suspension to new culture vessels.6. Incubate cultures at 37°C.Subcultivation Ratio: 1:3. Medium Renewal: Every 2 to 3 days.
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Complete growth medium 95%; DMSO, 5%
Storage: Store at -80 °C for less than 2 weeks. Store in vapor phase of liquid nitrogen for >2 weeks.
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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