Protocol for measuring in vivo, ex vivo, and in vitro glucose uptake in brown adipocytes

Here, we present a protocol for assessing in vivo glucose uptake in brown adipose tissue (BAT) from mice using 2-deoxy-D-[1,2=-³H]glucose. We describe steps for reagent and animal preparation, tracer administration, and tissue processing to quantify radiolabeled glucose uptake and its conversion to glucose-6-phosphate. We then detail procedures for conducting glucose uptake assays in freshly isolated BAT and cultured brown adipocytes, enabling assessment under a range of experimental conditions. For complete details on the use and execution of this protocol, please refer to Park et al.¹.