Selective binding of ethylene and ethylene action inhibitor in nanoporous materials can provide new strategies for harvest preservation technologies. In this work, we investigate the process of ethylene and 1-methylcyclopropene (1-MCP), also known as ethylene action inhibitor, binding on HKUST-1, a metal–organic framework (MOF) with open copper sites. For the first time, in situ infrared spectroscopy combined with theoretical simulations was used to obtain the three-dimensional structure of ethylene and 1-MCP bound to Cu-sites of HKUST-1. Time-resolved infrared spectra were collected to selectively determine the rates of ethylene desorption from Cu-sites and estimate its binding energies which were also compared with theoretical results. Not only a successful investigation of ethylene binding sites in HKUST-1 is presented but also a methodology that can be extended to study ethylene/1-MCP storage and release in porous materials with open metal sites which is relevant to the harvest preservation technology.