Preparation and Identification of Cardiac Myofibrils from Whole Heart Samples.
Preparation and Identification of Cardiac Myofibrils from Whole Heart Samples.
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DOI：
10.1007/978-1-0716-1480-8_2
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发表时间：
2021-01-01
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影响因子：
--
通讯作者：
Tong, Carl W
中科院分区：
文献类型：
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作者：
Creed, Heidi A;Tong, Carl W
小鼠模型被广泛研究并被很好地用于研究心肌病，因为这种模式生物的遗传相关性以及越来越多的需要确定治疗靶点。用于全心的心肌原纤维制剂已被证明是确定心力衰竭基本分子机制的一种非常有价值的体外方法。下面描述的技术持续产生完整的心肌肌原纤维，可用于后续技术，如蛋白质印迹、免疫荧光和质谱学。在这里，我们描述了一种优化从小鼠整个组织样本中分离和获得心肌原纤维的方法，并为随后的质谱分析做准备。这种方法允许快速视觉识别多个心肌肌原纤维蛋白。
Mouse models are extensively studied and well-used to study cardiomyopathies due to the genetic relevance of this model organism and increasing need to identify therapeutic targets. Cardiac myofibril preparation for whole hearts has proved to be an invaluable in vitro method for determining the fundamental molecular mechanisms in heart failure. The technique described below consistently yields intact cardiac myofibrils, which can be used in subsequent techniques such as western blotting, immunofluorescence, and mass spectrometry. Here, we describe a method to optimize the separation and yield of cardiac myofibrils from murine whole tissue samples and preparation for subsequent mass spectrometry. This method allows for quick visual identification of multiple cardiac myofibril proteins.