VisULite MAX ECL Western Blotting Substrate

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Detection of GAPDH in HeLa Cell Llysates Visualized with VisULite MAX^™ ECL Western Blotting Substrate Using X-ray Film or CCD Imaging System. A two-fold dilution series of HeLa whole cell lysate was prepared and loaded at 625, 313, 156, 78, 39, 19, 10 and 5 ng per lane. Samples were transferred onto PVDF membrane and probed with 0.1 µ/mL of Mouse Anti-Human/Mouse/Rat GAPDH Monoclonal Antibody (R&D Systems^®, Catalog # MAB5718) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (R&D Systems^®, Catalog # HAF018). VisULite MAXTM ECL substrate was applied for 1 minute, followed by exposure to a standard sensitivity film (left) or image capture using ProteinSimple^® FluorChem^™ M (right). Lanes are visualized on a 30 second exposure (film) or 1 minute exposure (imager).

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Detection of Recombinant Human CRP Visualized with VisULite MAX^™ ECL Western Blotting Substrate. A two-fold dilution series was prepared using rhCRP (R& Systems^®, Catalog # 1707-CR) and loaded at 50, 25, 12.5, 6.25, 3.13, 1.56, 0.78, and 0.39 pg per lane. Samples were transferred onto PVDF membrane and probed with 0.5 μg/mL of Sheep Anti-Human CRP (R& Systems^®, Catalog # AF1707) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (R& Systems^®, Catalog # HAF016). VisULite MAX^™ ECL substrate was applied for 1 minute, followed by exposure to a standard sensitivity film for 15 seconds.

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Signal Duration with VisULite MAX^™ ECL Western Blotting Substrate. A two-fold dilution series of HeLa whole cell lysate was prepared, loaded, transferred and probed with Mouse Anti-Human/Mouse/Rat GAPDH Monoclonal Antibody (R& Systems^®, Catalog # MAB5718). VisULite MAX^™ ECL substrate was applied for 1 minute to the PVDF membrane, followed by exposure to a standard sensitivity film for 1 minute at the indicated timepoints from 1 to 60 minutes after VisULite MAX^™ ECL substrate addition.