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免疫沉淀没有它无法实现

发布时间:2021-1-4 17:17:46      点击:

  今天要来讲讲离心机在免疫沉淀方面得重要性。

  JULEJIERANSEZHIMIANYICHENDIANJIANCHENGChIPYONGLAIYANJIUDNASHANGDETEDINGDANBAIZHI。

  ERRNAMIANYICHENDIANFA(RIP)HEChIPXIANGJIN,DANRNAMIANYICHENDIANFASHIYONGLAIYANJIUHUIYURNAJIEHEDEDANBAIZHI(RNA Binding ProteinImmunoprecipitation,RNAJIEHEDANBAIMIANYICHENDIAN),SHIYANJIUXIBAONEIRNAYUDANBAIJIEHEQINGKUANGDEJISHU,LIYONGZHENDUIMUBIAODANBAIDEKANGTIBAXIANGYINGDERNA-DANBAIFUHEWUCHENDIANXIALAI,TONGGUOLIXINJIFENLICHUNHUA,DUIJIEHEZAIFUHEWUSHANGDERNA JINXINGRT-PCRYANZHENGHUOCEXUFENXI,LEJIEZHUANLUHOUDIAOKONGWANGLUODONGTAIGUOCHENG,SHIYANJIUZHUANLUHOUDIAOKONGWANGLUODONGTAIGUOCHENGDEYOULIGONGJU,NENGBANGZHUWOMENFAXIANmiRNADEDIAOJIEBADIAN。

  RIPZHEIZHONGXINXINGDEJISHUYUNYONGZHENDUIMUBIAODANBAIDEKANGTIBAXIANGYINGDERNA-DANBAIFUHEWUCHENDIANXIALAI,RANHOUJINGGUOFENLICHUNHUAJIUKEYIDUIJIEHEZAIFUHEWUSHANGDERNAJINXINGFENXI。RIPKEYIKANCHENGSHIPUBIANSHIYONGDERANSEZHIMIANYICHENDIANChIPJISHUDELEISIYINGYONG,DANYOUYUYANJIUDUIXIANGSHIRNA-DANBAIFUHEWUERBUSHIDNA-DANBAIFUHEWU,RIPSHIYANDEYOUHUATIAOJIANYUChIPSHIYANBUTAIXIANGTONG(RUFUHEWUBUXUYAOGUDING,RIPFANYINGTIXIZHONGDESHIJIHEKANGTIJUEDUIBUNENGHANYOURNAMEI,KANGTIXUJINGRIPSHIYANYANZHENGDENGDENG)。

  RIPJISHUXIAYOUJIEHEmicroarrayJISHUBEICHENGWEIRIP-Chip,BANGZHUWOMENGENGGAOTONGLIANGDILEJIEAIZHENGYIJIQITAJIBINGZHENGTISHUIPINGDERNABIANHUA,JIEHEDERNAXULIETONGGUOmicroarray(RIP-Chip),DINGLIANGRT-PCRHUOGAOTONGLIANGCEXU(RIP-Seq)FANGFALAIJIANDING。

  MIANYICHENDIANSHILIYONGKANGTITEYIXINGFANYINGCHUNHUAFUJIMUDEDANBAIDEYIZHONGFANGFA。KANGTIYUXIBAOLIEJIEYEHUOBIAODASHANGQINGZHONGXIANGYINGDEDANBAIJIEHEHOU,ZAIYUDANBAIA/G(ProteinA/G)HUOERKANGOULIANDEagaroseHUOSepharoseZHUZIFUYU,TONGGUOLIXINDEDAOZHUZI-DANBAIA/GHUOERKANG-KANGTI-MUDEDANBAIFUHEWU,CHENDIANJINGGUOXIDIHOU,ZHONGXUANYUDIANYONGSHANGYANGHUANCHONGYE,ZHUFEI5-10min,ZAIGAOWENJIHAIYUANJIDEZUOYONGXIA,KANGYUANYUKANGTIJIELI,TONGGUOLIXINJILIXINSHOUJISHANGQING,SHANGQINGZHONGBAOKUOKANGTI、MUDEDANBAIHESHAOLIANGDEZADANBAI。

  MIANYIGONGCHENDIANSHIYIKANGTIHEKANGYUANZHIJIANDEZHUANYIXINGZUOYONGWEIJICHUDEYONGYUYANJIUDANBAIZHIXIANGHUZUOYONGDEJINGDIANFANGFA,SHIQUEDINGLIANGZHONGDANBAIZHIZAIWANZHENGXIBAONEISHENGLIXINGXIANGHUZUOYONGDEYOUXIAOFANGFA。

  MIANYIGONGCHENDIANSHIYANDAZHILIUCHENGWEI:SHOUJIDANBAIYANGPIN—YOUERDANBAIKANGTICHENDIANYOUERDANBAI—SDS-PAGE FENLIDANBAI—WB JIANCESHIFOUCUNZAIBADANBAI。

TGL-25M.jpg

  CAOZUOBUZHOU

  1.YIBANQIBUSHIYONG2GE10cm2MIN,YONGYULENGDEPBSXIDIXIBAOSANCI,ZUIHOUYICIXIGANPBS;

  2.JIARUXIBAOLIEJIEYE(westernJIIPLIEJIEYE)LIEJIEXIBAO,SHIDANBAIZHONGNONGDUZAI1-7mg/ml;

  3.4°LIXIN10min,QUCHUXIBAOSUIPIAN;ZHUYIZHEILIXUYAOYONGDAOLENGDONGLIXINJI,YINWEIWENDUSHANGYOUYANGEYAOQIU。

  4.GENJUYANGPINLIANGJIARU0.5-2ugZUOYOUKANGTI,4℃,XUANZHUANGUOYE。

  5.QU10-30ul protein A/G DEbeadsYUEPGUANZHONG,QISHANGQING,SHIYONGPBSXIDIbeadsYICI。

  6.JIANGYANGPINJIARUEPGUAN,4DUCHUIZHIHUNYUN2-3h。(ZAIXIQUQIONGZHITANGZHUDESHIHOU,ZHUYIYAOYAOYUNGUANZI,RANGZHUZIHERONGYEHUNHEJUNYUN,TONGSHIHUANGQIANGTOUYAOJIANDIAOTOUZIZAIXIQU,YIMIANPOHUAIZHUZI),PBSXITUO;4DULIXIN,QISHANGQING,

  7.XITUOSANCI,ZHUOQINGYONGPBS/PBST/GAOYANRONGYEXIQIONGZHITANGZHU,ZUIHOUYICIXIGANXIDIYE。

  8.JIANGSHILIANGXIANGYINGKANGTIDEDUOTAIHUO0.1M PH2.8DEGANANSUANXISHIDAOXIBAOLIEJIEYEZHONG,HUNYUN,QUSHILIANGYUbeadsZHONGJINXINGXITUO,QUCHUQINGZHONGLIAN。RUOYONGGANANSUANHCL,YANGPINXUYAOYONGNaOHZHONGHE,GENJUC1V1=C2V2JISUANJIARUNaOHDEHANLIANG。

  9.TONGGUOLIXINJILIXINQUSHANGQING,ZHIYANG,PAOJIAO。

  YISHANGJIUSHIGUANYULIXINJIZAIMIANYICHENDIANFANGMIANDEJIANGJIE,XIANZAININZHIDAO“MIANYICHENDIANMEIYOUTAWUFASHIXIAN”ZHONGDE"TA"SHUODESHISHENMELEBA!


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