| Cat.No.: | CSC-RI0182 |
| Cell Line Description: | HEK293-HuP2RX2 cell line is a hypotriploid human cell line, which has been transfected with a human purinergic receptor P2X, ligand-gated ion channel, 2 (P2RX2) to allow stably express of the human P2RX2. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system. |
| Background: | The human P2RX2 gene encodes P2X2, an ionotropic ATP receptor that functions as an ATP-activated cation channel expressed in the CNS and dorsal root ganglion neurons. P2X2 channels are therapeutic targets in treatment of pain. |
| Growth Properties: | Adherent |
| Morphology: | Epithelial |
| Host Cell: | HEK293 |
| Cell Line Validation: | 1. Gene expression: qPCR experiments determined specific expression of human P2RX2.2. Protein expression: P2RX2 expression in this cell line has been validated by WB.3. Functional validation: FLIPR, QPatch HT. |
| Application: | Therapeutic Area: pain & inflammation. |
| Sub-type: | Ionotropic purinergic receptors |
| Propagation: | Complete growth medium: DMEM/Complete growth medium: Ham's F12; 10% FBS; 500 μg/mL G418Atmosphere: 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 1:6 to 1:10 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: | P2RX2 purinergic receptor P2X, ligand-gated ion channel, 2 [ Homo sapiens ] |
| Official Symbol: | P2RX2 |
| Synonyms: | P2RX2; purinergic receptor P2X, ligand-gated ion channel, 2; P2X purinoceptor 2; P2X2; ATP receptor; P2X Receptor, subunit 2; MGC129601; |
| Gene ID: | 22953 |
| mRNA Refseq: | NM_012226 |
| Protein Refseq: | NP_036358 |
| MIM: | 600844 |
| UniProt ID: | Q9UBL9 |
| Chromosome Location: | 12q24.33 |
| Pathway: | Calcium signaling pathway, organism-specific biosystem; Calcium signaling pathway, conserved biosystem; Neuroactive ligand-receptor interaction, organism-specific biosystem; Neuroactive ligand-receptor interaction, conserved biosystem; |
| Function: | ATP binding; extracellular ATP-gated cation channel activity; extracellular ATP-gated cation channel activity; identical protein binding; ligand-gated ion channel activity; purinergic nucleotide receptor activity; purinergic nucleotide receptor activity; receptor activity; |
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