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HEK293 Human NPFFR2 NFAT-Luc Cell Line
Cat. No： RQP71627
Size： 1 vial of frozen cells (>1E6 per vial in 1 mL)
Unit Price： Contact For Pricing
Description
Product Info
Biological Info
Assay Data
Cell Culture
This product is a HEK293 stable cell line that stably expresses tag-free full-length human NPFFR2 receptor. Upon agonist binding to human NPFFR2, the receptor activates Gαq, triggering PLC cleavage and the generation of IP3 and DAG, leading to intracellular calcium mobilization and NFAT-luciferase expression. Therefore, receptor activity can be easily quantified in live cells in a dose-response manner by measuring luminescence signal. The cell line has been functionally validated with positive agonists. It is suitable for HTS and receptor functional evaluation assays.
| Cat. No | RQP71627 |
| Product Name | HEK293 Human NPFFR2 NFAT-Luc Cell Line |
| Engineering method | plasmid and lentivirus |
| Culture Properties | Adherent |
| Stability | 32 passages (in-house test, that not means the cell line will be instable beyond the passages we tested.) |
| Mycoplasma Status | Negative |
| Culture Medium | DMEM+10% FBS+2 μg/ml Puromycin +5 μg/ml Blasticidin +200 μg/ml Hygromycin B |
| Freeze Medium | 90% FBS+10% DMSO |
| Storage Conditions | Liquid nitrogen immediately upon delivery |
| Transducer | Gi/Go |
| Application | Functional assay for NPFFR2 |
| Safety Precautions |
Wear lab coat, disposable gloves to avoid direct sample contact. For spills: apply disinfectant, allow full inactivation, then clean up. |
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
| Target Class | GPCR |
| Family | Neuropeptide FF receptors (NPFFR) |
| Sub Family | Class A（Rhodopsin） |
| Gene Name | NPFFR2 |
| Gene Aliases |
NPFF2;NPGPR;GPR74
|
| Gene ID | 10886 |
| Accession Number | NM_004885 |
| UniProt Number | Q9Y5X5 |
| Protein Name | Neuropeptide FF receptor 2 |
| Protein Aliases | G protein-coupled receptor 74 (GPR74) ；NPFF2 receptor |
| Target Species | Human |
| Host cell | HEK293 |
Figure 1. Dose Response of Agonists in NPFFR2 NFAT-Luc HEK293(C38)
Neuropeptide AF and Neuropeptide FF induce dose-dependent luminescence response, EC50 values obtained for NPFFR2 functional validation.
Figure 2. HTRF cAMP Assay with NPFFR2 NFAT-Luc HEK293 (C38)
Dose-dependent cAMP responses induced by Neuropeptide AF and Neuropeptide FF, EC50 calculated for NPFFR2 functional validation.
Cell Resuscitation
1)Rapidly thaw the frozen cells in a 37 °C water bath for approximately 60 seconds. Once thawed (which may take slightly less or more than 60 seconds), immediately transfer the cell suspension from the cryovial into a 15 mL centrifuge tube containing 10 mL of pre-warmed HEK293 Human NPFFR2 NFAT-Luc Cell Line complete culture medium.
2)Centrifuge cells at 1000 rpm for 5 min to remove medium, then resuspend cells in 5 mL of pre-warmed complete medium.
3)Transfer the cell suspension into a T25 culture flask and incubate at 37 °C with 5% CO₂.
4)After approximately 24–36 hours, replace the medium or passage the cells to remove non-adherent dead cells.
Subculturing procedure
1)When the cell density reaches the appropriate confluency for passaging, wash the cells with PBS, then add 1 mL trypsin to detach the cells. When more than 80% of the cells detach upon gently tapping the culture flask, add complete culture medium to terminate digestion. Gently pipette to obtain a single-cell suspension, transfer to a 15 mL centrifuge tube, and centrifuge at 1000 rpm for 5 minutes.
2)Discard supernatant after centrifugation. Resuspend cells in fresh medium to a single-cell suspension and transfer to a new culture flask for continued growth.
Cell Freezing
After trypsinization and centrifugation of cells from each T75 flask or 10 cm culture dish, discard the supernatant. Add 2 mL of cryopreservation medium (90% FBS + 10% DMSO), gently resuspend thoroughly, and aliquot into two cryovials. Immediately place the cryovials into a controlled-rate freezing container (e.g., Nalgene 5100-0001), fill with isopropanol to the indicated level, and store at −80 °C. After 24 hours, transfer the cryovials to liquid nitrogen for long-term storage.
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