Display omitted •Stable isotopes are increasingly used tool of biologists.•Researchers often assume that isotopic properties are thermally static.•Temperature increased incorporation rates via higher growth and catabolism rates.•Temperature decreases trophic fractionation.•Natural variation in temperature must be accounted for when using stable isotopes. Stable isotopes are valuable tools in physiological and ecological research, as they can be used to estimate diet, habitat use, and resource allocation. However, in most cases a priori knowledge of two key properties of stable isotopes is required, namely their rate of incorporation into the body (incorporation rate) and the change of isotope values between consumers and resources that arises during incorporation of the isotopes into the consumer’s tissues (trophic discrimination). Previous studies have quantified these properties across species and tissue types, but little is known about how they vary with temperature, a key driver of many biological rates and times. Here, we explored for the first time how temperature affects both carbon incorporation rate and trophic discrimination via growth rates, using the domestic cricket, Acheta domesticus. We raised crickets at 16 °C, 21 °C, and 26 °C and showed that temperature increased carbon isotope incorporation rate, which was driven by both an increased growth rate and catabolism at higher temperatures. Trophic discrimination of carbon isotopes decreased at higher temperatures, which we attributed to either lower activation energies needed to synthesize non-essential amino acids at higher temperatures or the increased utilization of available resources of consumers at higher temperatures. Our results demonstrate that temperature is a key driver of both carbon isotope incorporation rate and trophic discrimination, via mechanisms that likely persist across all ectotherms. Experiments to determine incorporation rates and trophic discrimination factors in ectotherms must include temperature as a major factor, and natural variation in temperature might have significant effects on these isotopic properties that then can affect inferences made from isotope values.