Human Recombinant 4-1BB Stable Reporter Cell Line
HT1080/4-1BB Stable Reporter Cell Line

Figure 1. FACS analysis of cell surface expression of human 4-1BB on HT-1080/4-1BB cells. The HT-1080/4-1BB cells (blue) and the negative control HT-1080 cells (red) were probed using anti-human 4-1BB antibody and Alexa Fluor® 647-conjugated goat anti-human IgG (H+L) secondary antibody (Cat. No. A-21445, Thermo Fisher).

HT1080/4-1BB Stable Reporter Cell Line

Figure 2. Dose response of IL-8 secretion by HT-1080/4-1BB cell upon agonist 4-1BBL treatment. d2 acceptor fluorophore-labeled IL-8 (Cat. No. 62HIL08PET; Revvity) and IL-8 in the supernatant of HT-1080/4-1BB cells competitively bind with Europium Cryptate-labeled anti-IL-8 antibody. The FRET signal decreases as the IL-8 concentration in the supernatant rises and was measured by plate reader (Pherastar, BMG). The EC50 of 4-1BBL on HT-1080/4-1BB cells was 0.30 μg/ml.

HT1080/4-1BB Stable Reporter Cell Line

Figure 1. FACS analysis of cell surface expression of human 4-1BB on HT-1080/4-1BB cells. The HT-1080/4-1BB cells (blue) and the negative control HT-1080 cells (red) were probed using anti-human 4-1BB antibody and Alexa Fluor® 647-conjugated goat anti-human IgG (H+L) secondary antibody (Cat. No. A-21445, Thermo Fisher).

HT1080/4-1BB Stable Reporter Cell Line

Figure 2. Dose response of IL-8 secretion by HT-1080/4-1BB cell upon agonist 4-1BBL treatment. d2 acceptor fluorophore-labeled IL-8 (Cat. No. 62HIL08PET; Revvity) and IL-8 in the supernatant of HT-1080/4-1BB cells competitively bind with Europium Cryptate-labeled anti-IL-8 antibody. The FRET signal decreases as the IL-8 concentration in the supernatant rises and was measured by plate reader (Pherastar, BMG). The EC50 of 4-1BBL on HT-1080/4-1BB cells was 0.30 μg/ml.

HT1080/4-1BB Stable Reporter Cell Line

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4-1BB is a member of the tumor necrosis factor (TNF) receptor family. Its alternative names are tumor necrosis factor receptor superfamily member 9 (TNFRSF9), CD137 and induced by lymphocyte activation (ILA). It is currently of interest to immunologists as a co-stimulatory immune checkpoint molecule._x000D_ _x000D_ 4-1BB can be expressed by activated T cells, but to a larger extent on CD8 than on CD4 T cells. In addition, CD137_x000D_ expression is found on dendritic cells, follicular dendritic cells, natural killer cells, granulocytes and cells of blood vessel walls at sites of inflammation._x000D_ _x000D_ The best characterized activity of 4-1BB is its costimulatory activity for activated T cells. Crosslinking of 4-1BB enhances T cell proliferation, IL-2 secretion, survival and cytolytic activity. Further, it can enhance immune activity to eliminate tumors in mice.
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Product Introduction
Product Description Recombinant HT-1080 cells stably overexpress human 4-1BB on the surface. HT-1080 cell line is derived from human fibrosarcoma biopsy. It is highly active in NF-kB signaling pathway which is necessary for immune responses. The immune response of HT1080 cells overexpressing 4-1BB can be demonstrated by the increased production of IL-8.
Culture Properties Adherent.
Stability Stability: Stable through more than 16 passages with no significant changes in assay performance or expression profile.
Size Two vials of frozen cells (>1×106 per vial in 1 mL).
Storage Store cells in liquid nitrogen immediately upon receipt. Thaw and recover cells within one year from the date received.

Culture Conditions
Culture Medium MEM, 10% FBS, 200 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies), 2 μg/ml Puromycin (Cat. No. A11138-03, Life Technologies)
Complete Growth Medium MEM, (Cat. No. 41090, Life Technologies) 10% FBS (Cat. No. 10099-141, Life Technologies)
Freeze Medium-DATA 95% complete growth medium, 5% DMSO (Cat. No. D2650, Sigma)

Examples
  • HT1080/4-1BB Stable Reporter Cell Line
    • HT1080/4-1BB Stable Reporter Cell Line

    Figure 1. FACS analysis of cell surface expression of human 4-1BB on HT-1080/4-1BB cells. The HT-1080/4-1BB cells (blue) and the negative control HT-1080 cells (red) were probed using anti-human 4-1BB antibody and Alexa Fluor® 647-conjugated goat anti-human IgG (H+L) secondary antibody (Cat. No. A-21445, Thermo Fisher).

  • HT1080/4-1BB Stable Reporter Cell Line
    • HT1080/4-1BB Stable Reporter Cell Line

    Figure 2. Dose response of IL-8 secretion by HT-1080/4-1BB cell upon agonist 4-1BBL treatment. d2 acceptor fluorophore-labeled IL-8 (Cat. No. 62HIL08PET; Revvity) and IL-8 in the supernatant of HT-1080/4-1BB cells competitively bind with Europium Cryptate-labeled anti-IL-8 antibody. The FRET signal decreases as the IL-8 concentration in the supernatant rises and was measured by plate reader (Pherastar, BMG). The EC50 of 4-1BBL on HT-1080/4-1BB cells was 0.30 μg/ml.

  • HT1080/4-1BB Stable Reporter Cell Line
    • HT1080/4-1BB Stable Reporter Cell Line

    Figure 1. FACS analysis of cell surface expression of human 4-1BB on HT-1080/4-1BB cells. The HT-1080/4-1BB cells (blue) and the negative control HT-1080 cells (red) were probed using anti-human 4-1BB antibody and Alexa Fluor® 647-conjugated goat anti-human IgG (H+L) secondary antibody (Cat. No. A-21445, Thermo Fisher).

  • HT1080/4-1BB Stable Reporter Cell Line
    • HT1080/4-1BB Stable Reporter Cell Line

    Figure 2. Dose response of IL-8 secretion by HT-1080/4-1BB cell upon agonist 4-1BBL treatment. d2 acceptor fluorophore-labeled IL-8 (Cat. No. 62HIL08PET; Revvity) and IL-8 in the supernatant of HT-1080/4-1BB cells competitively bind with Europium Cryptate-labeled anti-IL-8 antibody. The FRET signal decreases as the IL-8 concentration in the supernatant rises and was measured by plate reader (Pherastar, BMG). The EC50 of 4-1BBL on HT-1080/4-1BB cells was 0.30 μg/ml.


For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.