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如何从脑浆组织中提取外泌体√
发布时间:2018-12-04   点击次数:216次

WAIMITISHIYOUXIBAONEIDUOPAOTIYUXIBAOMORONGHEHOU,SHIFANGDAOXIBAOWAIDEYIZHONGZHIJINGYUE 30 ~ 150 nm DEMOXINGNANGPAO。JIHUSUOYOUDEXIBAODOUHUICHANSHENGWAIMITI,BEIFENMICHUDEWAIMITIHUIJINRUGEZHONGTIYE,TONGGUOXUNHUANXITONGDAODAQITAXIBAOYUZUZHI,CHANSHENGYUANCHENGDIAOKONGZUOYONG。

越来越多的研究表明神经系统中的生理功能和病理功能与外泌体密切相关,例如:参与神经发育和神经元活动,参与神经退行性疾病调控等。
 

BENWENZHONGZHENGLILEYIZHONGKEYICONGNAOJIANGZUZHIZHONGFENLIHETIQUWAIMITIDECAOZUOBUZHOU,GONGDAJIACANKAO!

 


相关试剂:

彩搜网1,Hibernate E Solution(BrainBits, Springfield)

彩搜网2,MUGUADANBAIMEI papain(Worthington)

彩搜网3,Umibio Exosome Isolation Kit

4,PBS

 

相关设备:

1,YUNJIANGQI

2,WANGZHUANGGUOLVQI 40-μm

3,ZHUSHEQIGUOLVQI 0.2μm

4,GAOSULIXINJI

5,WOXUANZHENDANGQI

 

操作步骤:

一、脑浆液化处理:

1,JIANGXINXIANHUOXIANQIANLENGDONGDEXIAOSHUBANNAOQIEKAI,JIARU Hibernate E RONGYE (3 mL /BANNAO);

2,JIARU 20 DANWEI/ mL DEMUGUADANBAIMEI,ZAI 37℃ SHUIYU 15 FENZHONG;

彩搜网3,JIARUDENGTIJIYULENG Hibernate E HOU,QINGQINGYUNJIANG;YUNJIANGGUOCHENGZHONGZAIBINGSHANGJINXING;

4,NAOYUNJIANGTONGGUO 40-μm WANGZHUANGGUOLVQIGUOLV;

5,LVYEZAITONGGUO 0.2-μm GUOLVQIGUOLV,SHOUJILVYE;

 

二、液化脑浆预处理;

彩搜网1,LIXINQUSUIPIAN:JIANGYEHUANAOJIANGYEZHUANYIZHILIXINGUANZHONG,YU 4℃ YI 3000 g LIXIN 10 min,QUCHULVYEZHONGDESUIPIAN;LIXINHOUSHANGQINGYEZHUANYIDAOXINDELIXINGUANZHONG;

2,LIXINQUZAZHI:ZHUANYIHOUDESHANGQINGYEYU 4℃ YI 10000 g LIXIN 20 min,QUCHUYANGPINZHONGZAZHI,JIANGLIXINHOUDESHANGQINGYEZHUANYIZHIXINDELIXINGUANZHONG;

 

三、脑浆外泌体的提取;
 

TONGGUO Umibio Exosome Isolation Kit TIQUWAIMITIKELI。

彩搜网1,SHANGQINGYEYUCHULI:ZAIQUCHUZAZHIDELIXINSHANGQINGYEZHONGJIARU Exosomoe Concentration Solution(ECS SHIJI),JUTIDEJIARUJILIANGRUXIA:

样品名称

样品剂量

加入 PBS 溶液

加入 ECS 剂量

YEHUANAOJIANGYE

1 mL

4 mL

1 mL

彩搜网ZHU:QITAJILIANGGUIGEQINGGENJUBIAOZHONGDESHIJIYONGLIANGDENGBILIHUANSUAN

彩搜网2,RONGYEHUNHE:JIARU ECS SHIJIHOUJIANGLIXINGUANGAIJIN,TONGGUOWOXUANZHENDANGQIHUNYUN 1 min,ZAIFANGZHIYU 2℃ ZHI 8℃ JINGZHI 2 h;

彩搜网3,CHENDIANWAIMITI:QUCHUZHUANGYOUHUNHEYEDELIXINGUANYU 4℃ YI 10000 g LIXIN 60 min,QISHANGQING,CHENDIANZHONGFUHANWAIMITIKELI;(ZHU:JINKENENGXIJINGSHANGQINGYE)

彩搜网4,WAIMITIZHONGXUAN:QU 100μL 1×PBS JUNYUNCHUIDALIXINCHENDIANWU,DAIQIJUNYUNXUANFUZAI PBS ZHONGHOU,JIANGZHONGXUANYEZHUANYIZHIXINDE 1.5 mL LIXINGUANZHONG;

5,SHOUHUOWAIMITIKELI:JIANGHANYOUZHONGXUANYEDE 1.5 mL LIXINGUANYU 4℃ YI 12000 g LIXIN 2 min,BAOLIUSHANGQINGYE,GAISHANGQINGYEZHONGFUHANWAIMITIKELI。(ZHU:RUOLIXINCHENDIANWUROUYANKEJIAN,ZAILIXINYICI)

6, CHUNHUAWAIMITI:JIANGSHOUHUODEWAIMITIKELICUPINZHUANRU Exosomoe Purafication Filter(EPF ZHU)SHANGSHIZHONG,YU 4℃ YI 3000 g LIXIN 10 min,LIXINHOUSHOUJI EPF ZHUGUANDIDEYETI,CIYETIJIWEICHUNHUAHOUDEWAIMITIKELI;

彩搜网7,WAIMITIDEBAOCUN:CHUNHUAHOUDEWAIMITIYIBAOCUNYU-80℃ DIWENBINGXIANGZHONG,YIBEIHOUJISHIYANSHIYONG。

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