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It also directly stimulates mast cell degranulation and promotes the release of pruritogens including histamine and prostaglandins

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Western blot analysis involved preparing whole cell lysates using RIPA lysis buffer, quantifying protein concentration via BCA assay, followed by SDS-PAGE, transfer to nitrocellulose membranes, incubation with primary and HRP-conjugated secondary antibodies, and detection using an ECL solution [1]