The TCO-PEG5000-NHS ester is a heterobifunctional linker containing a TCO portion for inverse electron-demand Diels-Alder cycloaddition reactions and an NHS-activated ester.The TCO will react with the tetrazine via an inverse electron-demand Diels-Alder cycloaddition reaction to form a stabilized dihydropyridazinium bond. Extremely fast kinetics and selectivity enable the coupling of two low-abundance biopolymers in aqueous or other complex chemical environments. The excellent selectivity and biocompatibility of this bioorthogonal reaction allows complementary mates to react with each other in functionally rich biological systems, in some cases living organisms. Thus, tetrazine TCO linkages are used in fluorescence imaging, drug delivery, PET and SPECT imaging, and radionuclide therapy.