CHO-K1 Human OX1R Cell Line

CHO-K1 Human OX1R Cell Line

Cat. No: RQP71402

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 CHO-K1 stable cell line that stably expresses tag-free full-length human OX1R receptor. Upon agonist binding to human OX1R, the receptor activates Gαq, triggering PLC cleavage and the generation of IP3 and DAG, 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 or calcium flux. The cell line has been functionally validated with positive agonists. It is suitable for HTS and receptor functional evaluation assays.

Cat. No RQP71402
Product Name CHO-K1 Human OX1R 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 F12K+10% FBS+5 ug/ml puromycin
Freeze Medium 90% FBS+10% DMSO
Storage Conditions Liquid nitrogen immediately upon delivery
Transducer Gq
Application Functional assay for OX1R
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 Hypocretin receptors
Sub Family Class A(Rhodopsin)
Gene Name OX1R
Gene Aliases OXR1;ORXR1;HCRTR1
Gene ID 3061
Accession Number NM_001525
UniProt Number O43613
Protein Name Orexin/Hypocretin receptor type 1
Protein Aliases Hypocretin receptor type 1 ;Orexin receptor type 1 (Ox-1-R; Ox1-R; Ox1R)
Target Species Human
Host cell CHO-K1

HTRF IP-One assay dose response curve of Orexin A in OX1R CHO-K1(C10) cells, EC50 = 0.95 nM

Figure 1. HTRF IP-One Assay with OX1R CHO-K1(C10)

Orexin A shows potent agonist activity with EC50 = 0.95 nM. The cell line passes functional validation and is available for OX1R drug screening.

 

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 Human OX1R 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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