Crosslinking receptor-specific probes for electron microscopy

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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436525, 436526, 436501, 424 3, 424 71, 435 6, G01N 3353

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active

048792209

ABSTRACT:
A method and probes are disclosed for the localization of functional receptors by electron microscopy (EM). Electron dense probes are directed to receptors by means of specific ligand-receptor interactions and subsequently anchored covalently to target cellular material. A particular probe is a complex between colloidal gold and dextran derivatized with diaminohexane and containing photo-activatable nitroarylazido groups. Ligands with a specific affinity for certain receptors can be covalently attached to the complex to provide receptor-specific markers. For example, ouabain can be covalently attached to the dextran matrix to produce a probe specific for functional units of Na,K-ATPase. Covalent attachment of the bound probes at or near their respective receptors greatly enhances retention of probe during washing and standard procedures of fixation and dehydration in preparation for EM.

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