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yyc12596新蟲(chóng) (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(lái)的。 但是據(jù)說(shuō)ultracentrafuge也可以。對(duì)于超聲離心不熟悉，查查資料看看吧。 |
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yyc12596
新蟲(chóng) (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(shí)
- 蟲(chóng)號(hào): 2034994
- 注冊(cè): 2012-09-28
- 性別: MM
- 專業(yè): 白血病
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microvesicle的提�。� 1. Transfer equal volume of the supernatant (collected in step 10 in Subheading 3.1 ) to 3 ml fresh ultracentrifuge tubes ( see Note 8 ). 2. Mark the upper side of each ultracentrifuge tube with a waterproof mark. 3. Put ultracentrifuge tubes in the fi xed-angle rotor of ultracentrifuge with tweezers and make the waterproof mark face up. 4. Centrifuge for 2 h at 100,000 × g , 4 °C ( see Note 9 ). 5. Carefully take each ultracentrifuge tube out with tweezer ( see Note 10 ). 6. Note the position of microvesicle pellet and remove the supernatant completely (do not discard it) without disturbing the pellet ( see Note 11 ). 7. Collect the supernatant in a 50-ml polypropylene centrifuge tubes for later use. 8. Resuspend the pellet in each tube serially in 1 ml of PBS using pipetman ( see Note 12 ). 9. Pool the resuspended pellets from all the tubes containing urine from one subject in a single centrifuge tube. 10. Add appropriate volume of PBS if the volume of the collected pellet solution is less than three-quarters of the whole volume of the tube. Note the fi nal volume. 11. Make the waterproof mark on the side of the tube that contains sample. 12. Add equal volume of PBS to one other ultracentrifuge tube. It is used as a balancing tube. 13. Symmetrically put the two tubes in the fi xed-angle rotor of ultracentrifuge with tweezers and make the mark of the tube with sample face up. 14. Centrifuge for 2 h at 100,000 × g , 4 °C. 15. Carefully take each ultracentrifuge tube out with tweezers. 16. Note the position of microvesicle pellet and remove the supernatant completely without disturbing the pellet ( see Note 13 ). 17. Add 300 μ l of guanidine thiocyanate containing lysis buffer to the microvesicle pellet ( see Note 14 ). 18. Store microvesicle at −80 °C and avoid repeated freezing and thawing. |
9樓2014-08-05 00:07:55
yyc12596
新蟲(chóng) (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(shí)
- 蟲(chóng)號(hào): 2034994
- 注冊(cè): 2012-09-28
- 性別: MM
- 專業(yè): 白血病
★ ★ ★
youlinglyw: 金幣+3, 贈(zèng)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(zèng)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(shí)驗(yàn)室的Ultracentrifuge是Beckman Coulter的Optima™ L-80 XP。 我之前沒(méi)用過(guò)。 公司對(duì)于儀器的說(shuō)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(chóng) (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(shí)
- 蟲(chóng)號(hào): 2034994
- 注冊(cè): 2012-09-28
- 性別: MM
- 專業(yè): 白血病
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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(chóng) (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(shí)
- 蟲(chóng)號(hào): 2034994
- 注冊(cè): 2012-09-28
- 性別: MM
- 專業(yè): 白血病
4樓2014-08-04 23:35:51
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