描述The Click-iT® Lipid Peroxidation Imaging Kit - Alexa Fluor® 488 leverages copper-catalyzed click chemistry and the linoleamide alkyne (LAA) reagent (alkyne-modified linoleic acid) for detection of lipid peroxidation-derived protein modifications in fixed cells. Linoleic acid is the most abundant polyunsaturated fatty acid found in mammals and its lipid peroxidation products likely account for the majority of lipid-derived protein carbonyls.The Click-iT® Lipid Peroxidation Imaging Kit is: • Optimized for detection of lipid peroxidation in cells using click chemistry and Alexa Fluor® 488 azide • Multiplexible and compatible with traditional fluorescence microscopy, high content screening (HCS), and flow cytometry • Provided as a complete kit sufficient for five 96-well plates or 100 coverslips How It Works The Click-iT® Lipid Peroxidation Imaging Kit contains cumene hydroperoxide as a positive control for induction of lipid peroxidation and all of the components for the click reaction, including LAA, and Alexa Fluor® 488 azide. LAA can be incubated with cells, where it incorporates into cellular membranes. Upon lipid peroxidation, LAA is oxidized and produces 9- and 13- hydroperoxy-octadecadienoic acid (HPODE). These hydroperoxides decompose to multiple α,β-unsaturated aldehydes, which readily modify proteins at nucleophilic side chains. These alkyne-containing modified proteins can be subsequently detected using Click-iT® chemistry and multiplexed with other probes appropriate for fixed cells. This assay is amenable to detection by traditional fluorescence microscopy, high content screening (HCS), and flow cytometry. Available in Two Formats The Click-iT® Lipid Peroxidation Imaging Kit - Alexa Fluor ® 488 contains the components you need to detect lipid peroxidation in cells, including positive control compound. Alternatively, Click-iT® LAA (linoleamide alkyne) is a stand-alone reagent provided as a five pack set of single-use vials for maximum flexibility in assays for lipid peroxidation. This reagent may be combined with a variety of azide-modified detection reagents and other reagents in the Click-iT® tool box. For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. |
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