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Panoply™ Human CCR8 Over-expressing Stable Cell Line

Panoply™ Human CCR8 Over-expressing Stable Cell Line

Cat.No. :  CSC-SC002678 Host Cell:  HEK293 (CHO and other cell types are also available)

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Gene Informationn

Cat. No. CSC-SC002678
Description Using Creative Biogene's proprietary lentiviral vectors, we subclone the target gene into lentivector, generate the lentivirus particles, sequentially infect the cell line HEK293 (other cell types are also available according to your requirements), and select the clones constantly expressing target gene at high level.
Gene CCR8
Gene Species Homo sapiens (Human)
Host Cell HEK293 (CHO and other cell types are also available)
Stability Validated for at least 10 passages
Application

1. Gene expression studies

2. Signaling pathway research

3. Drug screening and toxicology

4. Disease research

Quality Control Negative for bacteria, yeast, fungi and mycoplasma.
Size Form 2 × 10^6 cells / vial
Shipping Dry Ice
Storage Liquid nitrogen
Revival Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media.
Gene Name
Gene Symbol
Synonyms
Gene ID
UniProt ID
Chromosome Location
Function
Pathway
MIM
Mycoplasma Negative
Format One frozen vial containing millions of cells
Storage Liquid nitrogen
Safety Considerations

The following safety precautions should be observed.

1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.

2. No eating, drinking or smoking while handling the stable line.

3. Wash hands after handling the stable line and before leaving the lab.

4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.

5. All waste should be considered hazardous.

6. Dispose of all liquid waste after each experiment and treat with bleach.

Ship Dry ice
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The C-C motif chemokine receptor 8 (CCR8) belongs to the class A G protein-coupled receptor (GPCR). CCR8 interacts with the human-specific chemokine ligand CCL1/I-309, which is produced by a variety of cells, including tumor-associated macrophages and regulatory T cells (Tregs). CCR8 is highly expressed in Tregs and T helper 2 (TH2) cells recruited to sites of inflammation and has been implicated in allergies, asthma, and cancer progression. CCR8+ Tregs are considered important regulators of the immunosuppressive tumor microenvironment (TME); therefore, the development of sensitive monoclonal antibodies (mAbs) targeting CCR8 has been highly anticipated. Here, researchers developed a specific mAb against human CCR8 (hCCR8) for flow cytometric detection in cell-based immunity and screening (CBIS) approaches. Flow cytometry revealed that the established anti-human CCR8 monoclonal antibody, C8Mab-21 (murine IgM, κ type), reacted with hCCR8-overexpressing Chinese Hamster Ovary-K1 cells (CHO/hCCR8), TALL-1 cells (acute T-lymphoblastic leukemia), CCRF-HSB2 cells (human T-lymphoblastic leukemia), and natural killer cells, which express endogenous hCCR8 by flow cytometry. Furthermore, C8Mab-21 exhibited moderate affinity for both CHO/hCCR8 and TALL-1 cells, with dissociation constants of 6.5×10⁻⁸ M and 2.0×10⁻⁸ M, respectively. C8Mab-21, constructed using the CBIS method, can serve as an effective tool for flow cytometric analysis of hCCR8-related biological responses.

Flow cytometric analysis of CHO-K1, hCCR8-overexpressing CHO-K1 cells, TALL-1, CCRF-HSB2, and NK cells was performed using C8Mab-21 and commercially available anti-human CD198 (CCR8) mAbs (clones S19017D and L263G8). C8Mab-21, S19017D, and L263G8 recognized hCCR8-overexpressing CHO-K1 cells in a dose-dependent manner (Figure 1A). Even at a mAb concentration of 20 μg/mL, C8Mab-21 and L263G8 did not react with parental CHOK1 cells. S19017D showed a slight reaction with CHO-K1 cells at 20 μg/mL and 2 μg/mL (Figure 1B). Regarding endogenously hCCR8-expressing cells, C8Mab-21 recognized TALL-1, CCRF-HSB2, and NK cells at a concentration of 2 μg/mL or 20 μg/mL. S19017D also responded to TALL-1 and CCRF-HSB2 in a dose-dependent manner even at a concentration of 0.02 μg/mL. However, S19017D failed to recognize NK cells even at a concentration of 20 μg/mL. L263G8 also responded to TALL-1 and CCRF-HSB2 even at a concentration of 0.02 μg/mL and also to NK cells at mAb concentrations of 2 μg/mL or higher. C8Mab-21 detected both exogenously and endogenously expressed naïve hCCR8 by flow cytometry.

Figure 1. Flow cytometric analysis of anti-hCCR8 mAbs against CHO/hCCR8 and CHO-K1.Figure 1. Flow cytometric analysis of anti-hCCR8 mAbs against CHO/hCCR8 and CHO-K1. (Tanaka T, et al. 2024)

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