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.