求使用蔗糖梯度密度离心分离和蔗糖垫超离exosome的文章

hzangs

RT
求一篇使用蔗糖梯度密度离心分离exosome的文章，再求一篇使用蔗糖垫加超离分离exosome的文章。  文章领域不限，只求其method里的方法是这个即可。 谢谢

子非鱼

Sonia A. Melo, et al. Glypican-1 identifies cancer exosomes and detects early pancreatic cancer. nature,2015.这篇里的斑竹看看？
-------"Sucrose gradient. Sucrose density gradients were performed to purify exosomes. Exosomes were resuspended in 2 ml of HEPES/sucrose stock solution (2.5 M sucrose, 20 mM HEPES/NaOH solution, pH 7.4). The exosomes suspension was overlaid with a linear sucrose gradient (2.0–0.25 M sucrose, 20 mM HEPES/ NaOH, pH 7.4) in a SW41 tube (Beckman). The gradients were ultracentrifuged for 16 h at 10,000g at 4uC. Gradient fractions of 1 ml were collected from top to bottom and densities of each fractions were evaluated using a refractometer. Next, the exosomes pellets were washed in PBS followed by a second step of ultracentrifugation at 150,000g at 4uC for 2 h. Exosomes pellets were resuspended in
Laemmli buffer and/or PBS for further immunoblot and flow cytometry analysis."

hzangs

子非鱼 发表于 2016-1-9 19:52
Sonia A. Melo, et al. Glypican-1 identifies cancer exosomes and detects early pancreatic cancer. nat ...

谢谢

wjy

Before exosome isolation, Huh 7-ET or infected Huh7.5.1 cells
were cultured for 48 h. Exosomes were isolated as described before (27). In
题目：Exosome-mediated transmission of hepatitis C virus between human hepatoma Huh7.5 cells
杂志PNAS
作者：Vedashree Ramakrishnaiaha, Christine Thumannb,c, Isabel Fofanab,c, Francois Habersetzerb,c, Qiuwei Pand,
Petra E. de Ruitera, Rob Willemsene, Jeroen A. A. Demmersf, Victor Stalin Rajg, Guido Jensterh, Jaap Kwekkeboomd,
Hugo W. Tilanusa, Bart L. Haagmansg, Thomas F. Baumertb,c, and Luc J. W. van der Laana,1

brief, cell culture supernatants were subjected to successive centrifugations
of 3,042 × g (20 min) and 10,000 × g (30 min). Exosomes were then pelleted
at 64,000 × g for 110 min, using an SW28 rotor (Beckman Coulter Instruments). Exosome pellets were resuspended in 0.32 M sucrose and centrifuged at 100,000 × g for 1 h (SW60Ti rotor; Beckman Coulter Instruments).
The exosome pellet was then resuspended in PBS

hzangs

wjy 发表于 2016-1-10 19:59
Before exosome isolation, Huh 7-ET or infected Huh7.5.1 cells
were cultured for 48 h. Exosomes were  ...

谢谢

wangjoseph

Palma J et al. Nucl. Acids Res. 2012, Vol. 40, No. 18 9125–9138
MicroRNAs are exported from malignant cells in customized particles

Royo et al., Plos one, 2013, 8(7): e68693. doi:10.1371/journal.pone.0068693
Transcriptome of Extracellular Vesicles Released by Hepatocytes