细胞膜蛋白分离方法(超速离心法)

1.seed cells for 24:00:00 in a 10 cm dish at a density of 6 x 106 cells per plate

2.wash cells with 1X PBS for 2 times

3.scrape cells of the dish and harvest in 1 ml pf PBS + PI buffer

4.1000 x g, 00:15:00

5.resuspend cells in 500 µl PBS + PI buffer

6.sonicate 2 times at 30Hz for 15 ON-OFF intervals of 10 s each (1 s pulse on/off).

7.340000 x g, 4°C, 00:15:00

collect supernatant as cytosolic fraction

8.was pellet 3 times with PBS + PI buffer

9.340000 x g, 4°C, 00:15:00

10.disolve pellet in 500 µL 1x PBS +PI+ 1% Trtiton

11.340000 rpm, 4°C, 00:15:00

collect supernatant as membranous fraction

如若转载,请注明出处:https://www.ouq.net/cellmemisolated.html

(0)
打赏 微信打赏,为服务器增加50M流量 微信打赏,为服务器增加50M流量 支付宝打赏,为服务器增加50M流量 支付宝打赏,为服务器增加50M流量
上一篇 03/04/2022 18:47
下一篇 03/05/2022 01:10

相关推荐