CHO-K1 Human CRF1R Cell Line
Cat. No： RQP71434
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 CHO-K1 stable cell line that stably expresses tag-free full-length human CRF1R receptor. Upon agonist binding to human CRF1R, the receptor activates Gs, which stimulates adenylate cyclase and increases intracellular cAMP, leading to activated PKA/CREB signaling and HPA axis response. Therefore, receptor activity can be easily quantified in live cells in a dose-response manner by measuring cAMP. The cell line has been functionally validated with positive agonist Urocortin. It is suitable for HTS and receptor functional evaluation assays.
| Cat. No | RQP71434 |
| Product Name | CHO-K1 Human CRF1R 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 μg/ml puromycin |
| Freeze Medium | 90% FBS+10% DMSO |
| Storage Conditions | Liquid nitrogen immediately upon delivery |
| Transducer | Gs |
| Application | Functional assay for CRF1R |
| 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 | Corticotropin releasing hormone receptors |
| Sub Family | Class B1（Secretin） |
| Gene Name | CRF1R |
| Gene Aliases | CRHR1;CRHR;CRF-R;CRF1 |
| Gene ID | 1394 |
| Accession Number | NM_004382 |
| UniProt Number | P34998 |
| Protein Name | CRF-R-1; CRF-R1; CRFR-1 |
| Protein Aliases | CRH-R-1; CRH-R1 |
| Target Species | Human |
| Host cell | CHO-K1 |
Figure 1. HTRF cAMP Assay with CRF1R CHO-K1 Cell(C27)
Urocortin inhibits cAMP signal in a dose-dependent manner, with EC₅₀ = 7.5 pM. The cell line is suitable for CRF1R pharmacological 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 CRF1R 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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