Seoane-Collazo Patricia, López Miguel
NeurObesity Group, Department of Physiology, CIMUS, University of Santiago de Compostela-Instituto de Investigación Sanitaria, Santiago de Compostela, Spain.
CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), Santiago de Compostela, Spain.
Methods Mol Biol. 2018;1732:433-448. doi: 10.1007/978-1-4939-7598-3_27.
Hypothalamic AMPK plays a key role in the control of energy homeostasis by regulating energy intake and energy expenditure, particularly modulating brown adipose tissue (BAT) thermogenesis. The function of AMPK can be assayed by analyzing its phosphorylated protein levels in tissues, since AMPK is activated when it is phosphorylated at Thr-172. Here, we describe a method to obtain hypothalamic (nuclei-specific) protein extracts and the suitable conditions to assay AMPK phosphorylation by Western blotting.
下丘脑 AMPK 通过调节能量摄入和能量消耗，特别是调节棕色脂肪组织（BAT）产热，在能量平衡控制中发挥关键作用。AMPK 的功能可以通过分析其在组织中的磷酸化蛋白水平来检测，因为当 AMPK 在苏氨酸 172 位点被磷酸化时它就会被激活。在这里，我们描述一种获取下丘脑（细胞核特异性）蛋白提取物的方法以及通过蛋白质印迹法检测 AMPK 磷酸化的合适条件。
