CHO-K1 Human IL6RA/IL6ST Dimerization Cell

CHO-K1 Human IL6RA/IL6ST Dimerization Cell

Cat. No: RQP74258

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 reporter cell line that stably expresses tag-free full-length human IL6RA (interleukin-6 receptor subunit alpha) together with IL6ST (interleukin-6 signal transducer, also known as GP130). Upon IL6 binding, IL6RA forms a complex with IL6ST and induces IL6ST dimerization, which activates the JAK-STAT pathway, mainly through STAT1 and STAT3, as well as the RAS-dependent MAPK cascade, driving reporter signal output. Receptor dimerization and downstream pathway activation can therefore be quantified in live cells in a dose-response manner by measuring the reporter signal.

Signal pathway diagram of hIL6-induced IL6Rα-GP130 dimerization detected by split luciferase reporter assayFigure 1. Schematic diagram of IL6RA /IL6ST Dimerization CHO-K1 Cell Model

Cat. No RQP74258
Product Name CHO-K1 Human IL6RA/IL6ST Dimerization Cell
Product Type Reporter Cell
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 blasticidin+5 μg/ml puromycin
Freeze Medium 90% FBS+10% DMSO
Storage Conditions Liquid nitrogen immediately upon delivery
Application Functional(Report Gene) Assay
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.

Classification Cytokine&Growth Factor
Family type I cytokine receptor family. Type 3 subfamily
Gene Name IL6R
Gene Aliases CD126;IL-6R;IL-1Ra;IL6RA;gp80
Gene ID 3570
Accession Number NM_000565.4
UniProt Number P08887
Protein Name IL-6 receptor subunit alpha; IL-6R subunit alpha; IL-6R-alpha; IL-6RA
Protein Aliases IL-6R 1;Membrane glycoprotein 80 (gp80)
Family-2 type I cytokine receptor family. Type 2 subfamily
Gene Name-2 IL6ST
Gene Aliases-2 GP130;CD130;sGP130;IL-6RB
Gene ID-2 3572
Accession Number-2 NM_002184.4
UniProt Number-2 P40189
Protein Name-2 IL-6 receptor subunit beta; IL-6R subunit beta; IL-6R-beta; IL-6RB
Protein Aliases-2 CDw130;Interleukin-6 signal transducer;gp130
Target Species Human
Host cell CHO-K1

Flow cytometry histograms confirming IL6RA and IL6ST expression in IL6RA/IL6ST dimerization CHO-K1 (C39) cells versus parental CHO-K1 control cells stained with corresponding antibodies

Figure 1. Recombinant IL6RA/IL6ST Dimerization CHO-K1 constitutively expressing IL6RA/IL6ST

The test group exhibits distinctly higher PE fluorescence signals compared with the parental CHO-K1 control, confirming successful co-expression of both target proteins for IL-6 induced dimerization assay.

Dose-response curve of human IL6 in IL6RA/IL6ST dimerization CHO-K1 (C39) reporter cell line, EC50 = 4.76 ng/ml, measured by split luciferase luminescence (RLU)

Figure 2. Dose response of  Human IL6 in IL6RA/IL6ST Dimerization CHO-K1 Cell(C39)

The luminescence signal rises with increasing hIL6 concentration, with an EC50 of 4.76 ng/ml, demonstrating reliable functional activity of the cell model for IL6 pathway 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 IL6RA/IL6ST Dimerization Cell 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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