Flagship species are among key marketing tools used by conservation organizations to motivate public support, but are often selected in an ad hoc, rather than systematic, manner. Furthermore, it is unclear whether selected flagship species do motivate public support. This paper describes a multi-method exploratory study, carried out in Switzerland, which aimed to determine the selection criteria for flagship species and measure whether a species selected according to these criteria was able to motivate support. Fourteen representatives of international, regional and local conservation organizations were interviewed and the selection criteria for their flagship species were identified. A charismatic species (the great spotted woodpecker) that meets these criteria and an apparently less charismatic species (the clover stem weevil) were selected as treatments in a quantitative experiment with 900 respondents. Using conjoint analysis, it was found that both charismatic and uncharismatic species have the ability to positively influence public preferences for habitat variables that encourage biodiversity in urban landscapes. These results may be used by conservation organizations to assist in the selection of flagship species, and in particular for flagship species that are intended to perform a specific conservation function.