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Immobilon FL


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  • Indoleamine 2,3-dioxygenase mediates the antiviral effect of gamma interferon against hepatitis B virus in human hepatocyte-derived cells. 21084489

    Alpha interferon (IFN-α) is an approved medication for chronic hepatitis B. Gamma interferon (IFN-γ) is a key mediator of host innate and adaptive antiviral immunity against hepatitis B virus (HBV) infection in vivo. In an effort to elucidate the antiviral mechanism of these cytokines, 37 IFN-stimulated genes (ISGs), which are highly inducible in hepatocytes, were tested for their ability to inhibit HBV replication upon overexpression in human hepatoma cells. One ISG candidate, indoleamine 2,3-dioxygenase (IDO), an IFN-γ-induced enzyme catalyzing tryptophan degradation, efficiently reduced the level of intracellular HBV DNA without altering the steady-state level of viral RNA. Furthermore, expression of an enzymatically inactive IDO mutant did not inhibit HBV replication, and tryptophan supplementation in culture completely restored HBV replication in IDO-expressing cells, indicating that the antiviral effect elicited by IDO is mediated by tryptophan deprivation. Interestingly, IDO-mediated tryptophan deprivation preferentially inhibited viral protein translation and genome replication but did not significantly alter global cellular protein synthesis. Finally, tryptophan supplementation was able to completely restore HBV replication in IFN-γ- but not IFN-α-treated cells, which strongly argues that IDO is the primary mediator of IFN-γ-elicited antiviral response against HBV in human hepatocyte-derived cells.
    Document Type:
    Reference
    Product Catalog Number:
    MAB1501R
    Product Catalog Name:
    Anti-Actin Antibody,clone C4

  • Do I need to rinse the Immobilon-FL blotting membrane with water prior to blocking step, after wetti…

    It is not necessary to rinse the membrane with water. However, carry-over of methanol from the wetting step can reduce binding of blocking reagents to the membrane.
    Document Type:
    FAQ
    Product Catalog Number:
    C3117
    Product Catalog Name:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • How long does the fluorescent signal stay on the developed blot?…

    The fluorescent signal from fluorescence-tagged antibodies will remain stable on the membrane for several months, or longer, when stored protected from light. The membranes may be stored dry or in PBS at 4ºC. The fluorescent signal from chemi-fluorescent substrates does not remain for long so blots using these substrates need to be scanned immediately.
    Document Type:
    FAQ
    Product Catalog Number:
    C3117
    Product Catalog Name:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • Is Immobilon-FL better than Nitrocellulose membrane for Fluorescence detection?…

    Yes. Background fluorescence of Immobilon-FL is typically 2-5X lower than that of nitrocellulose, thus improving signal-to-noise ratio (sensitivity). Nitrocellulose membranes have other disadvantages. If allowed to dry out, they become brittle, tend to fracture and are difficult to handle. They are not recommended for stripping and re-probing. Nitrocellulose blots need to be scanned as soon as possible after detection, as diffusion of signal on a wet membrane may also occur.
    Document Type:
    FAQ
    Product Catalog Number:
    C3117
    Product Catalog Name:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • Do I need to dry Immobilon-FL blots before scanning?…

    Drying the blot may enhance signal strength. NOTE: Do not wrap the blot in plastic/saran wrap while scanning. If you do need to wrap the membrane, Mylar should be used.
    Document Type:
    FAQ
    Product Catalog Number:
    C3117
    Product Catalog Name:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • What is the level of detection sensitivity when using Immobilon-FL?…

    Immobilon-FL exhibits the lowest background fluorescence of any blotting membrane thus improving the signal-to-noise ratio (sensitivity) of any fluorescent dye. The fluorescence signal is dependent on the fluorescent dye used, thus the ultimate sensitivity of detection is determined by the dye itself. With Immobilon-FL the sensitivity is not limited by the membrane, e.g., detection of proteins in the low picogram level has been observed with QDot® Nanocrystals fluorescent-tagged antibodies on Immobilon-FL.
    Document Type:
    FAQ
    Product Catalog Number:
    C3117
    Product Catalog Name:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • Can Immobilon-FL be used with Millipore’s Rapid Immunodetection Method?…

    Yes. Immobilon-FL can be used with the Rapid Immunodetection Method. However slightly higher background may be observed.
    Document Type:
    FAQ
    Product Catalog Number:
    C3117
    Product Catalog Name:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • What fluorescent dyes are recommended to use with Immobilon-FL?…

    Immobilon-FL membrane exhibits very low background fluorescence over a broad range of wavelengths and can be used with all visible and infrared fluorescent dyes. Generally, dyes with larger Stokes shift (greater separation between absorption and emission wavelengths) are preferred in western blotting applications using fluorescence-based detection. NOTE: To prevent photo-bleaching, protect the membrane from light during secondary antibody incubations and washes until the membrane is ready to be scanned.
    Document Type:
    FAQ
    Product Catalog Number:
    C3117
    Product Catalog Name:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper