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yyc12596新虫 (著名写手)
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microvesicle ultracentrafuge 已有1人参与
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microvesicle获取。 文献中有用AnnexinV磁珠拉下来的。 但是据说ultracentrafuge也可以。对于超声离心不熟悉，查查资料看看吧。 |
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yyc12596
新虫 (著名写手)
- 应助: 10 (幼儿园)
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文献1的离心方法： Ultracentrifugation and microvesicle isolation Microvesicle-free supernatant was generated by ultracentrifugation (2.5 h, 100,000 g) and subsequently used for ELISA analysis. To isolate microvesicles, cell supernatant from senescent cells was subjected to serial ultracentrifugation. Briefly, conditioned, serum-free cell supernatant was first cleared from dead cells and debris by low-speed centrifugation (10 min., 1000 g) and afterward spun for 30 min at 10,000 g to precipitate larger vesicles. In order to enrich for exosomes, the resulting supernatant was filtered through a 0.2 m filter prior to ultracentrifugation at 100,000 g for 2.5 h (Optima LE-80K, Type 70 Ti Rotor; Beckman Coulter, Krefeld, Germany). The microvesicle pellet was washed with PBS, transferred to cell lysis buffer, and used for Western blotting. |
5楼2014-08-04 23:43:28
yyc12596
新虫 (著名写手)
- 应助: 10 (幼儿园)
- 金币: 10847.3
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- 虫号: 2034994
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- 专业: 白血病
★ ★ ★
youlinglyw: 金币+3, 赠人玫瑰手有余香，生物科学有你更精彩！ 2014-08-05 09:42:24
youlinglyw: 金币+3, 赠人玫瑰手有余香，生物科学有你更精彩！ 2014-08-05 09:42:24
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实验室的Ultracentrifuge是Beckman Coulter的Optima™ L-80 XP。 我之前没用过。 公司对于仪器的说明网址： https://www.beckmancoulter.com/w ... /2/392049///0/1//0/ |
2楼2014-08-04 23:31:26
yyc12596
新虫 (著名写手)
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- 专业: 白血病
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配套的软件： Optima's easy-to-use eXPert Software gives your lab an edge by making it possible to quickly design and optimize separation protocols using a combination of rotors and tubes to simulate results — all before performing separations. A unique slide-bar feature allows you to view the separation at earlier run times to better suit the isolation of individual components. Up to three components can be modeled using the rate-zonal protocol. Then save the simulated run and download it with a single click of the mouse when you're ready to begin your run. Optima eXPert Software's advanced intelligence delivers greater productivity and faster turnarounds for your lab. The following demonstrations illustrate just how easy it is to optimize efficiency with Optima eXPert Software. |
3楼2014-08-04 23:34:31
yyc12596
新虫 (著名写手)
- 应助: 10 (幼儿园)
- 金币: 10847.3
- 散金: 2230
- 红花: 34
- 帖子: 2270
- 在线: 355.1小时
- 虫号: 2034994
- 注册: 2012-09-28
- 性别: MM
- 专业: 白血病
4楼2014-08-04 23:35:51