HEK293 Human NK3R CRE-Luc Cell Line

HEK293 Human NK3R CRE-Luc Cell Line

Cat. No: RQP71625

Size: 1 vial of frozen cells (>1E6 per vial in 1 mL)

Unit Price: Contact For Pricing

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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 NK3R receptor with CRE-Luc reporter. Upon agonist binding to human NK3R, the receptor activates Gαq and Gαs, triggering PLC cleavage and the generation of IP3, leading to intracellular calcium mobilization, IP1 accumulation and cAMP elevation. 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 RQP71625
Product Name HEK293 Human NK3R CRE-Luc Cell Line
Engineering method plasmid transfection
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+200 μg/ml Hygromycin B
Freeze Medium 90% FBS+10% DMSO
Storage Conditions Liquid nitrogen immediately upon delivery
Transducer Gs
Application Functional assay for NK3R
Safety Precautions

Wear lab coat, disposable gloves to avoid direct sample contact.

For spills: apply disinfectant, allow full inactivation, then clean up.

Seal samples properly during transportation to prevent leakage and biological contamination.

 

 

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 NK3R
Gene Aliases TACR3;NKR;TAC3R
Gene ID 6870
Accession Number NM_001059
UniProt Number P29371
Protein Name NKR
Protein Aliases NK-3 receptor (NK-3R);Neurokinin B receptor;Tachykinin receptor 3
Target Species Human
Host cell HEK293

Flow cytometry histogram showing high NK3R expression in HEK293 stable cell line versus parental cell control.
Figure 1.  Recombinant NK3R CRE-Luc HEK293 stably expressing NK3R

Dose-response curves of multiple agonists tested on NK3R CRE-Luc HEK293(C36) cell line, with calculated EC50 values, demonstrating NK3R receptor pharmacological activity.
Figure 2. Dose Response of Agonists in NK3R CRE-Luc HEK293(C36)

Multiple agonists were applied to characterize NK3R activity, and EC50 values were determined.
HTRF cAMP dose-response curves of NK3R CRE-Luc HEK293(C36) with multiple agonists. EC50 values were determined to evaluate NK3R receptor pharmacological activity.
Figure 3. HTRF cAMP Assay with NK3R CRE-Luc HEK293(C36)

Dose-response profiles of multiple agonists, EC50 values calculated to characterize NK3R activity.

HTRF IP-One dose-response curves of NK3R CRE-Luc HEK293(C36) tested with multiple agonists. EC50 values obtained to verify NK3R pharmacological function.
Figure 4. HTRF IP-One Assay with NK3R CRE-Luc HEK293(C36)

Dose-response curves of various agonists, EC50 values calculated to characterize NK3R receptor activity.

 

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 NK3R 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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