Gonzales Patricia A, Zhou Hua, Pisitkun Trairak, Wang Nam Sun, Star Robert A, Knepper Mark A, Yuen Peter S T
Laboratory of Kidney and Electrolyte Metabolism, NHLBI, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2010;641:89-99. doi: 10.1007/978-1-60761-711-2_6.
Exosomes represent an important and readily isolated subset of the urinary proteome that has the potential to shed much insight on the health status of the kidney. Each segment of the nephron sheds exosomes into the urine. Exosomes are rich in potential biomarkers, especially membrane proteins such as transporters and receptors that may be up- or downregulated during disease states. Two differential centrifugation methods are available for simple purification of exosomes: one uses ultracentrifugation, and the other uses a nanomembrane concentrator. Validation methods include western blots of pan-exosome markers and segment-specific exosome markers, and negative staining electron microscopy.
外泌体是尿蛋白质组中一个重要且易于分离的子集,有潜力为肾脏健康状况提供诸多见解。肾单位的每个节段都会向外泌体中释放外泌体。外泌体富含潜在的生物标志物,尤其是膜蛋白,如转运蛋白和受体,它们在疾病状态下可能会上调或下调。有两种差速离心方法可用于简单纯化外泌体:一种使用超速离心,另一种使用纳米膜浓缩器。验证方法包括全外泌体标志物和节段特异性外泌体标志物的蛋白质免疫印迹,以及负染色电子显微镜检查。