HEK293 Human NK1R CRE-Luc Cell Line
Cat. No： RQP71615
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 NK1R receptor with CRE-Luc reporter. Upon agonist binding to human NK1R, the receptor activates Gαs and Gαq, triggering adenylate cyclase for cAMP elevation and PLC cleavage to generate IP3, leading to intracellular calcium mobilization and IP1 accumulation. Therefore, receptor activity can be easily quantified in live cells in a dose-response manner by measuring IP1, cAMP or luciferase signal. The cell line has been functionally validated with positive agonists. It is suitable for HTS and receptor functional evaluation assays.
| Cat. No | RQP71615 |
| Product Name | HEK293 Human NK1R CRE-Luc Cell Line |
| Engineering method | plasmid transfection |
| Culture Properties | Adherent |
| Stability | 32passages (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 + 200μg/ml Hygromycin B |
| Freeze Medium | 90% FBS+10% DMSO |
| Storage Conditions | Liquid nitrogen immediately upon delivery |
| Transducer | Gq |
| Application | Functional assay for NK1R |
| 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 | Tachykinin receptors |
| Sub Family | Class A（Rhodopsin） |
| Gene Name | NK1R |
| Gene Aliases | TACR1;TAC1R;SPR;NKIR |
| Gene ID | 6869 |
| Accession Number | NM_001058 |
| UniProt Number | P25103 |
| Protein Name | Substance-P receptor |
| Protein Aliases | SPR;NK-1 receptor (NK-1R);Tachykinin receptor 1 |
| Target Species | Human |
| Host cell | HEK293 |
Figure 1.Dose Response of Agonists in NK1R CRE-Luc HEK293(C1)
Substance P TFA exhibits potent activation (EC50 =11.9nM), Neurokinin A TFA shows weaker activity (EC50=71.6nM), while EB1002 has no obvious activation (EC50>5000nM).
Figure 2. HTRF cAMP Assay with NK1R CRE-Luc HEK293(C1) in 500 μM IBMX
Substance P TFA (EC50 = 2.15 nM) and Neurokinin A TFA (EC50 = 4.67 nM) potently activate NK1R to elevate cAMP. EB1002 exhibits negligible activity with EC50 > 5000 nM.
Figure 3. HTRF IP-One Assay with NK1R CRE-Luc HEK293(C1)
Neurokinin A TFA (EC50 = 0.27 nM) and Substance P TFA (EC50 = 0.56 nM) strongly trigger IP1 accumulation. EB1002 activates NK1R with weaker potency (EC50 = 253.7 nM).
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 NK1R CRE-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.