Chapter title |
Isolation of Extracellular Vesicles in Saliva Using Density Gradient Ultracentrifugation
|
---|---|
Chapter number | 27 |
Book title |
Extracellular Vesicles
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7253-1_27 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7251-7, 978-1-4939-7253-1
|
Authors |
Kazuya Iwai, Satoshi Yamamoto, Mitsutaka Yoshida, Kiyotaka Shiba |
Abstract |
This chapter describes a method for isolating human salivary extracellular vesicles (EVs) using density gradient ultracentrifugation. Standard protocols established for isolation of EVs from blood or a conditioned medium of cultured cells do not work for whole saliva, due to its viscosity. Therefore, procedures including a pretreatment step and utilizing iodixanol as a gradient material enable EVs to be concentrated to a 1.1 g/ml density. This protocol is compatible with both swing and angle rotors. By employing an angle rotor, which enables high g-force, the centrifugation time was reduced to 4 h from the 17 h required when using a swing rotor. |
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