Covalent cholesterol conjugates have been prepared by isoxazole-generating [3+2] nitrile oxide alkyne cycloaddition (NOAC) chemistry. Steroidal building blocks functionalised with either an alkyne or a nitrile oxide precursor were evaluated. The reaction has been demonstrated for the tethering to cholesterol of groups capable of bioreporting, and for the formation of cholesterol- biomolecule conjugates. Nitrile oxide alkyne cycloaddition (NOAC) chemistry - a tool for attaching cholesterol to biomolecules or groups capable of bioreporting. Copyright Â© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.