Biotinylated reagents are molecules chemically modified to include biotin, a small vitamin (B7) that binds with exceptionally high affinity and specificity to avidin, streptavidin, or neutravidin proteins—a non-covalent interaction (Kd ≈ 10⁻¹⁵ M) that forms the foundation of many sensitive detection systems in molecular biology. When paired with enzyme-conjugated avidin or biotinylated molecules—often labeled with enzymes like horseradish peroxidase (HRP) or alkaline phosphatase (AP)—this system enables powerful signal amplification for detecting specific targets. Upon binding a biotinylated probe to its target (e.g., antibody, lectin, or nucleic acid), the enzyme-linked partner catalyzes a substrate to generate a visible signal—colorimetric, fluorescent, or chemiluminescent—making these reagents indispensable in techniques like immunohistochemistry, flow cytometry, glycan analysis, ELISA, and Western blotting.
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