Detecting low-abundance biomarkers in Immunoassays
Ultra Beacons dramatically boost the performance of immunoassays. In this sandwich immunoassay, the analyte is captured between a capture antibody and a detection antibody. We compared a conventional dye-labeled detection antibody (7 dyes) with the same antibody labeled using Ultra Beacons carrying 1, 30, 80, or 140 dyes. For each variant, we measured the lower limit of detection (LoD) and used its inverse as a proxy for assay sensitivity. As the dye load of the Ultra Beacon increases, so does the signal intensity and assay sensitivity. Replacing the standard label with an Ultra Beacon improved immunoassay performance by 23-fold, simply by swapping one reagent.
Brighter tissue staining for multiplexed immunofluorescence
Ultra Beacons enhance CD8α visualization in paraffin-embedded human tonsil tissue. We used a low concentration of biotinylated anti-CD8α antibody to model low-expressed targes. Under identical imaging settings, SA-UB-AF647 at just 20 nM produced stronger fluorescence and clearer staining than conventional SA-AF647 at 200 nM - demonstrating improved visualization at a tenfold lower detection-reagent concentration.
Detecting low-abundance targets in Flow Cytometry
Ultra Beacons enable highly sensitive staining of rare antigens in flow cytometry. In this experiment, a cancer cell line expressing low-abundance targets was stained either with a conventional commercially available antibody or with the same antibody labeled using an Ultra Beacon. Conventional antibodies show limited separation between the stained sample and the isotype control. In contrast, Ultra Beacons amplify the fluorescence signal, producing a clear separation and improving population resolution. As a result, Ultra Beacons substantially increase the staining index (the difference between the mean fluorescence intensities of the stained sample and the isotype control, divided by the standard deviation of the isotype control), enabling more accurate detection of rare cell-surface markers.
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