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yyc12596新蟲 (著名寫手)
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[交流]
microvesicle ultracentrafuge 已有1人參與
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microvesicle獲取。 文獻(xiàn)中有用AnnexinV磁珠拉下來的。 但是據(jù)說ultracentrafuge也可以。對于超聲離心不熟悉，查查資料看看吧。 |
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yyc12596
新蟲 (著名寫手)
- 應(yīng)助: 10 (幼兒園)
- 金幣: 10847.3
- 散金: 2230
- 紅花: 34
- 帖子: 2270
- 在線: 355.1小時
- 蟲號: 2034994
- 注冊: 2012-09-28
- 性別: MM
- 專業(yè): 白血病
|
文獻(xiàn)1的離心方法： 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樓2014-08-04 23:43:28
yyc12596
新蟲 (著名寫手)
- 應(yīng)助: 10 (幼兒園)
- 金幣: 10847.3
- 散金: 2230
- 紅花: 34
- 帖子: 2270
- 在線: 355.1小時
- 蟲號: 2034994
- 注冊: 2012-09-28
- 性別: MM
- 專業(yè): 白血病
★ ★ ★
youlinglyw: 金幣+3, 贈人玫瑰手有余香，生物科學(xué)有你更精彩！ 2014-08-05 09:42:24
youlinglyw: 金幣+3, 贈人玫瑰手有余香，生物科學(xué)有你更精彩！ 2014-08-05 09:42:24
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實驗室的Ultracentrifuge是Beckman Coulter的Optima™ L-80 XP。 我之前沒用過。 公司對于儀器的說明網(wǎng)址： https://www.beckmancoulter.com/w ... /2/392049///0/1//0/ |
2樓2014-08-04 23:31:26
yyc12596
新蟲 (著名寫手)
- 應(yīng)助: 10 (幼兒園)
- 金幣: 10847.3
- 散金: 2230
- 紅花: 34
- 帖子: 2270
- 在線: 355.1小時
- 蟲號: 2034994
- 注冊: 2012-09-28
- 性別: MM
- 專業(yè): 白血病
|
配套的軟件： 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樓2014-08-04 23:34:31
yyc12596
新蟲 (著名寫手)
- 應(yīng)助: 10 (幼兒園)
- 金幣: 10847.3
- 散金: 2230
- 紅花: 34
- 帖子: 2270
- 在線: 355.1小時
- 蟲號: 2034994
- 注冊: 2012-09-28
- 性別: MM
- 專業(yè): 白血病
4樓2014-08-04 23:35:51
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