CHO-K1 Rat CTR/RAMP1(AMY1) CRE-Luc Cell Line

CHO-K1 Rat CTR/RAMP1(AMY1) CRE-Luc Cell Line

Cat. No: RQP71657

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 CRE-Luc reporter CHO-K1 cell line that stably expresses tag-free full-length rat CTR together with RAMP1. Upon agonist binding to rat CTR-RAMP1 complex, the receptor activates Gαs, which stimulates adenylate cyclase and increases intracellular cAMP, triggering CRE-luciferase reporter expression. Therefore, receptor activity can be easily quantified in live cells in a dose-response manner by measuring luciferase signal. The cell line has been functionally validated with positive agonists. It is suitable for HTS and receptor functional evaluation assays.

Cat. No RQP71657
Product Name CHO-K1 Rat CTR/RAMP1(AMY1) CRE-Luc Cell Line
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 F12K + 10% FBS + 5 μg/ml Puromycin + 5 μg/ml Blasticidin + 600 μg/ml Hygromycin B
Freeze Medium 90% FBS+10% DMSO
Storage Conditions Liquid nitrogen immediately upon delivery
Transducer Gs
Application Functional assay for Rat CTR/RAMP1(AMY1R)
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
Sub Family Class B1 (Secretin)
Family-1 G-protein coupled receptor 2 family
Gene Name-1 Calcr
Gene Aliases-1 Clr, Ct-r
Gene ID-1 116506
Accession Number-1 NM_001034015
UniProt Number-1 P32214
Protein Name-1 Calcitonin receptor
Protein Name-1 Calcitonin receptor
Protein Aliases-1 CT-R, C1A/C1B, CTR
Family-2 RAMP family
Gene Name-2 Ramp1
Gene ID-2 29444
Accession Number-2 NM_012944
UniProt Number-2 P50451
Protein Name-2 Receptor activity-modifying protein 1
Protein Aliases-2 N/A
Target Species Rat
Host cell CHO-K1

 

  Dose response curve of agonists in Rat CTR/RAMP1 CRE-Luc CHO-K1 cell line, showing EC50 values: Rat Amylin 0.2141 nM, human Amylin 0.06564 nM, Cagrilintide 2.819 nM, Salmon Calcitonin 0.04711 nM, Eloralintide 8.24 nM
Figure 1. Dose Response of Agonists in Rat CTR/RAMP1(AMY1) CRE-Luc CHO-K1(C36) 

Salmon Calcitonin exhibits the highest potency with EC50 of 0.04711 nM, followed by human Amylin, rat Amylin, Cagrilintide and Eloralintide. The cell line enables reliable CRE-luciferase reporter detection for CTR/RAMP1(AMY1) target drug screening.

HTRF cAMP assay curve of Rat CTR/RAMP1 CRE-Luc CHO-K1 cell line, EC50 values: Rat Amylin 0.3559 nM, human Amylin 0.2596 nM, Cagrilintide 1.43 nM, Salmon Calcitonin 0.04849 nM, Eloralintide 3.065 nM
Figure 2. HTRF cAMP Assay with Rat CTR/RAMP1(AMY1) CRE-Luc CHO-K1(C36) 

Salmon Calcitonin shows the highest potency with EC50 of 0.04849 nM, followed by human Amylin, rat Amylin, Cagrilintide and Eloralintide. This cell line supports cAMP-based functional validation for CTR/RAMP1(AMY1) drug discovery.

 

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 CHO-K1 Rat CTR/RAMP1(AMY1) 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.

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