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yyc12596

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[交流] microvesicle ultracentrafuge 已有1人参与

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

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尿样准备:
1. Collect fresh urine in a 500 ml sterile plastic bottle ( see Note 2 ).
2. Transfer fresh urine to 50 ml polypropylene centrifuge tubes( see Note 3 ).
3. Centrifuge for 30 min at 3,000 × g , at 4 °C, using refrigerated centrifuge with swinging-bucket rotors.
4. Pipet off the supernatant, and dispense into 2-ml screw-cap Eppendorf ultracentrifuge tubes. Discard sediment and dispose the 50-ml polypropylene centrifuge tubes properly ( see Note 4 ).
5. Mark the cap of each ultracentrifuge tube with a waterproof mark.
6. Put the tubes in the fi xed-angle rotor of tabletop ultracentrifuge.
7. Orient the tubes in the rotor and make the waterproof mark face up ( see Note 5 ).
8. Close tabletop ultracentrifuge and centrifuge for 30 min at 13,000 × g , at 4 °C.
9. Take out the tube, note the position of cell debris, and carefully draw the supernatant with 1 ml syringe without disturbing the sediment ( see Note 6 ).
10. Inject the supernatant into 0.22 μ m fi lter and collect the supernatant with 50 ml polypropylene centrifuge tubes ( see Note 7 ).
8楼2014-08-05 00:06:08
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yyc12596

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★ ★ ★
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
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
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yyc12596

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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
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yyc12596

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设置时间:
Optima eXPert Software Demo 2:
Run Simulation -- Rate Zonal

Find out how to stop your run at the optimum separation time before the sample material reaches the tube bottom and becomes a pellet.
4楼2014-08-04 23:35:51
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