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Oil and gas has traditionally had spatially limited impacts in terrestrial systems, but new technologies are enabling unprecedented scales of development across large areas of North American rangelands, with unknown impacts for grassland biodiversity. Protected areas are considered by the conservation community to be one of the most effective means of mitigating for biodiversity loss. However, novel patterns and scales of oil and gas development represent the potential for new anthropogenic impacts to diminish their effectiveness. Our study identifies the extent of oil and gas development within protected areas and in the matrix in which they are embedded. We use proprietary and publicly available oil and gas datasets to establish the amount of oil and gas wells within protected areas in North America, and quantify the extent and magnitude of oil and gas development in the matrix surrounding protected areas. More than 25,000 wells were drilled in protected areas between 1900 and 2012, with the number of new oil and gas wells increasing over time. Increased development within protected areas is largely restricted to those established for sustainable resource use. However, high numbers of wells are found in proximity to all categories of protected areas. This development of the matrix surrounding conservation lands could have long lasting impacts on the ecosystem dynamics within these areas, as a formerly connected network of protected patches is transformed into isolated islands in an industrialized matrix. If protected areas are to continue to effectively conserve biodiversity, a multi-scale assessment of the effects of oil and gas development is necessary to advance conservation planning from local scales to biome levels. This provides an opportunity for rangeland ecologist to work with industry on land-use planning and restoration to avoid future loss of biodiversity in the face of large-scale conversions of rangelands.