Exosome Isolation from Plasma Samples

Plasma samples were pass through a 0.8 μm filter to remove additional cellular fragments and cell debris before exosome isolation (Millipore Millex). Exosomes were collected using the exoEasy Maxi Kit (Qiagen) [7 (link)]. A total of 10 ml buffer XWP were added and centrifuge at 5000g for 5 min to wash exosomes. We used 400 μL of Buffer XE (Qiagen) to dissolve the exosomes, centrifuged at 5000g for 5 min and then collected the eluate. All steps followed the manufacturer’s instructions. Purified exosomes were then stored at − 80 °C until use.

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Kuang M., Tao X., Peng Y., Zhang W., Pan Y., Cheng L., Yuan C., Zhao Y., Mao H., Zhuge L., Zhou Z., Chen H, & Sun Y. (2019). Proteomic analysis of plasma exosomes to differentiate malignant from benign pulmonary nodules. Clinical Proteomics, 16, 5.

Publication 2019

exoEasy Maxi Kit Buffer XE

Buffer Cell isolation Cellular Exosomes Plasma

Corresponding Organization :

Other organizations : Fudan University Shanghai Cancer Center, Center for Excellence in Molecular Cell Science, Shanghai Medical College of Fudan University, Second Military Medical University

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Variable analysis

independent variables

Exosome isolation method (Millipore Millex filter and exoEasy Maxi Kit)

dependent variables

Purified exosomes

control variables

Volume of buffer XWP (10 ml)
Centrifugation speed (5000g)
Centrifugation duration (5 min)
Volume of Buffer XE (400 μL)
Storage temperature (- 80 °C)

controls

Positive control: Not mentioned
Negative control: Not mentioned

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