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The International Union for Conservation of Nature (IUCN) Red List of Threatened Species, a species extinction risk assessment tool, has been guiding conservation efforts for over 5 decades. It is widely assumed to have been instrumental in preventing species from moving closer to extinction and driving recoveries. However, the impact of the IUCN Red List in guiding conservation has not been evaluated. We conducted, transcribed, and coded interviews with experts who use the IUCN Red List across a range of sectors to understand how the list is used in conservation. We developed a theory of change to illustrate how and why change is expected to occur along causal pathways contributing to the long-term goal of the IUCN Red List and an evaluation framework with indicators for measuring the impact of the IUCN Red List in generating scientific knowledge, raising awareness among stakeholders, designating priority conservation sites, allocating funding and resources, influencing development of legislation and policy, and guiding targeted conservation action (key themes). Red-list assessments were the primary input leading to outputs (scientific knowledge, raised awareness), outcomes (better informed priority setting, access to funding and resource availability, improved legislation and policy), and impact (implemented conservation action leading to positive change) that have resulted in achievement of IUCN Red List goals. To explore feasibility of attributing the difference made by the IUCN Red List across themes, we studied increased scientific knowledge, raised awareness, access to funding and resource allocation, and increased conservation activity. The feasibility exploration showed increased scientific knowledge over time identified through positive trends in publications referring to the IUCN Red List in the literature; raised awareness of the list following high IUCN activity identified by peaks in online search activity; an increased proportion of conservation funding bodies requesting IUCN Red List status in the application process; and, based on interviews with Amphibian Specialist Group members, red-list assessments were essential in connecting relevant stakeholders and ensuring conservation action. Although we identified the IUCN Red List as a vital tool in global conservation efforts, it was challenging to measure specific impacts because of its ubiquitous nature. We are the first to identify the influence of the IUCN Red List on conservation.