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Newborn Calf Serum

新生小牛血清

Company: Sigma-Aldrich
Catalog#: N4887
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Assessment of Brown Adipocyte Thermogenic Function by High-throughput Respirometry
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2015-11-05
[Abstract]  Brown adipose tissue (BAT) has the unique ability to dramatically increase mitochondrial uncoupled fuel oxidation for thermogenesis in response to adrenergic stimulation. A key parameter in assessing brown adipocyte thermogenic capacity is mitochondrial uncoupling as determined by respiration. Measuring mitochondrial oxygen consumption rate (OCR) therefore provides valuable information to study the regulation and dysregulation of fuel metabolism and energy expenditure. Adding measurements of mitochondrial membrane potential allows for more in-depth interpretation of the respirometry data. Here we provide protocols for measuring respiration in adherent intact and plasma membrane permeabilized brown adipocytes using the Seahorse XF Analyzer. In the protocol Part I, a combination of ... [摘要]  棕色脂肪组织(BAT)具有显着增加线粒体解偶联燃料氧化以响应肾上腺素刺激的热生成的独特能力。评估棕色脂肪细胞产热能力的关键参数是通过呼吸确定的线粒体解偶联。因此,测量线粒体氧消耗率(OCR)为研究燃料代谢和能量消耗的调节和失调提供了有价值的信息。添加线粒体膜电位的测量允许更加深入地解释呼吸数据。在这里我们提供使用Seahorse XF分析仪测量贴壁完整和质膜透性棕色脂肪细胞呼吸的协议。在方案部分I中,去甲肾上腺素和游离脂肪酸的组合用于诱导解偶联呼吸。然后使用ATP合酶抑制剂寡霉素,化学去偶联剂FCCP和复合物III抑制剂抗霉素A分别测量偶联的,最大的和非线粒体的氧消耗。在方案第II部分中,质膜用重组perfringolysin O透化,胆固醇依赖性细胞溶解素寡聚化成在质膜中专门的孔。这允许代谢物可用性的实验性控制,而不从天然细胞环境中分离线粒体。

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