WB检测

 

Western Blot(WB),JIDANBAIZHIYINJIFA(MIANYIYINJISHIYAN)。

 

TASHIFENZISHENGWUXUE、SHENGWUHUAXUEHEMIANYIYICHUANXUEZHONGCHANGYONGDEYIZHONGSHIYANFANGFA。QIJIBENYUANLISHITONGGUOTEYIXINGKANGTIDUININGJIAODIANYONGCHULIGUODEXIBAOHUOSHENGWUZUZHIYANGPINJINXINGZHESE。TONGGUOFENXIZHESEDEWEIZHIHEZHESESHENDUHUODETEDINGDANBAIZHIZAISUOFENXIDEXIBAOHUOZUZHIZHONGBIAODAQINGKUANGDEXINXI。

 

DANBAIZHIYINJIFASHIYOURUISHIMIXIEERFULEIDELIXISHENGWUYANJIUSUO(Friedrich Miescher Institute)DEHarry TowbinZAI1979NIANTICHUDE。ZAINIER·BONAITE(Neal Burnette)YU1981NIANSUOZHUDE《FENXISHENGWUHUAXUE》(Analytical Biochemistry)ZHONGSHOUCIBEICHENGWEIWestern Blot。DANBAIMIANYIYINJI(Western Blot)SHIJIANGDIANYONGFENLIHOUDEXIBAOHUOZUZHIZONGDANBAIZHICONGNINGJIAOZHUANYIDAOGUXIANGZHICHIWUNCMOHUOPVDFMOSHANG,RANHOUYONGTEYIXINGKANGTIJIANCEMOUTEDINGKANGYUANDEYIZHONGDANBAIZHIJIANCEJISHU,XIANYIGUANGFANYINGYONGYUJIYINZAIDANBAISHUIPINGDEBIAODAYANJIU、KANGTIHUOXINGJIANCEHEJIBINGZAOQIZHENDUANDENGDUOGEFANGMIAN。

 

 

原理

 

YUSouthern BlotHUONorthern BlotZAJIAOFANGFALEISI,DANWestern BlotFACAIYONGDESHIJUBINGXIXIANANNINGJIAODIANYONG,BEIJIANCEWUSHIDANBAIZHI,“TANZHEN”SHIKANGTI,“XIANSE”YONGBIAOJIDEERKANG。JINGGUOPAGE(JUBINGXIXIANANNINGJIAODIANYONG)FENLIDEDANBAIZHIYANGPIN,ZHUANYIDAOGUXIANGZAITI(LIRUXIAOSUANXIANWEISUBOMO)SHANG,GUXIANGZAITIYIFEIGONGJIAJIANXINGSHIXIFUDANBAIZHI,QIENENGBAOCHIDIANYONGFENLIDEDUOTAILEIXINGJIQISHENGWUXUEHUOXINGBUBIAN。YIGUXIANGZAITISHANGDEDANBAIZHIHUODUOTAIZUOWEIKANGYUAN,YUDUIYINGDEKANGTIQIMIANYIFANYING,ZAIYUMEIHUOTONGWEISUBIAOJIDEDIERKANGTIQIFANYING,JINGGUODIWUXIANSEHUOFANGSHEZIXIANYINGYIJIANCEDIANYONGFENLIDETEYIXINGMUDEJIYINBIAODADEDANBAICHENGFEN。GAIJISHUYEGUANGFANYINGYONGYUJIANCEDANBAISHUIPINGDEBIAODA。

 

 

【CAOZUOBUZHOU】

 

YI、DANBAIYANGPINZHUNBEI

SHOUJIDANBAIYANGBEN,JIARUDANBAISHANGYANGHUANCHONGYEHUNYUN,100℃JIARE5 min,CHONGFENBIANXINGDANBAI。

ER、DIANYONGYUZHUANMO

1,SDS-PAGENINGJIAOZHIBEI;

2,QUSHILIANGYANGBENSHANGYANG,XIANZAINONGSUOJIAOZHONGYONG100-120 VDIANYONGPAOJIAOZHIFENLIJIAO,HOUZENGJIADIANYADAO120-150 VDIANYONGZHIMUDEDANBAIYIJINGZUIDAHUADEFENLI;

3,DIANYONGJIESHU,JIANGFENLIJIAOZHUANYIZHIYULENGDEZHUANMOYEZHONG,200-250 mAHENGLIUZHUANMO2 h。

SAN、DANBAIZAJIAOJIXIANSE

1,XIMO:ZHUANMOJIESHUHOU,JIANGDANBAIMOFANGZHIDAOYUXIANZHUNBEIHAODEWesternXIDIYEZHONG,ZHIZHIXIMOYEXIJIN;

2,FENGBI:JIANGMOXIAOXINFANGRUFENGBIYEZHONG,YAOCHUANGSHANGQINGWEIYAOHUANG,SHIWENXIAFENGBI 1-1.5 GEXIAOSHI,HUO4℃ FENGBIGUOYE;

3,YIKANGFUYU:XIJINFENGBIYE,JIARUXISHIHAODEYIKANG,SHIWENYAOCHUANGSHANGFUYU1.5XIAOSHIHUOZHE4°CFUYUGUOYE;

4,ERKANGFUYU:JIANGMOZHUANYIZHIXISHIHAODEERKANG,SHIWENXIAZAJIAO1-2XIAOSHI。

5,ECHUAXUEFAGUANG,XIANYING,DINGYING,JIANGJIAOPIANJINXINGSAOMIAOHUOPAIZHAO。

 

 

实例展示

 

(Xu S , Wang P , Zhang J , et al. [J]. Molecular Cancer, 2019, 18)

 

BENYANJIUTONGGUOQUANZHUANLUZUCEXU,SHAIXUANBINGSUODINGZHANGLIANFEIBIANMARNA EGOTZUOWEIYANJIUBABIAO。TONGGUOSHIYANBIAOMINGEGOTNENGZENGJIARUXIANAIXIBAODUIZISHANCHUNDEMINGANXING,BINGQIEZAIJIZHISHANGCIJISUTONGGUOER-NRIP1-AP1SANYUANFUHETI,ZHUANLUYIZHIEGOTDEBIAODA,CONGERZENGJIAZHONGLIUXIBAOZISHISHUIPINGBINGTIGAODUIZISHANCHUNDEMINGANXING。

 

 

参考文献:

 

Dennis-Sykes, Carolyn A., William J. Miller, and William J. McAleer. "A quantitative Western Blot method for protein measurement." Journal of biological standardization 13.4 (1985): 309-314.

Rawlins, Mindy L., et al. "Evaluation of a western blot method for the detection of Yersinia antibodies: evidence of serological cross-reactivity between Yersinia outer membrane proteins and Borrelia burgdorferi." Clin. Diagn. Lab. Immunol. 12.11 (2005): 1269-1274.

Griffiths, Steven G., et al. "Comparison of western blot, direct fluorescent antibody and drop-plate culture methods for the detection ofRenibacterium salmoninarum in Atlantic salmon (Salmo salar L.)." Aquaculture 97.2-3 (1991): 117-129.

Liu, Chengyi, et al. "Roles of autophagy‑related genes Beclin‑1 and LC3 in the development and progression of prostate cancer and benign prostatic hyperplasia." Biomedical reports 1.6 (2013): 855-860.

 

 

 

 

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