亚博app官网包装和保存条件
SHIJIHEYU-20℃BAOCUN。Lysis Buffer -20℃KEBAOCUNYINIAN。FANYINGSHIJI(Luciferase Reaction ReagentHELuciferase Reaction Reagent II)JIARUDIWUHOU,FENZHUANG、BIGUANGBAOCUN;-70℃KEBAOCUNYINIAN,-20℃KEBAOCUNYIGEYUE;BIMIANFANFUDONGRONG。SHIJISHIYONGQIANXUPINGHENGZHISHIWEN。
实验原理
YINGHUOCHONGYINGGUANGSUMEI(firefly luciferase, fluc)HEHAISHENYINGGUANGSUMEI(renilla luciferase, rluc)KEFENBIECUIHUAYINGGUANGSU(luciferin)HEQIANGCHANGSU(coelenterazine)CHANSHENGSHENGWUYINGGUANGFANYING,FANYINGGUOCHENGRUXIATUSUOSHI。
ZAISHUANGYINGGUANGSUMEIFANYINGZHONG,SHOUXIANYONGLuciferase Reaction ReagentJIANCEfLucXINHAO;SUIHOU,ZHIJIEJIARULuciferase Reaction Reagent II,JIANGfLucDEXINHAOCUMIE,TONGSHICHANSHENGrLucXINHAO。GAIFANGFAJUYOUCAOZUOJIANDAN、JIANCEKUAIJIE、LINGMINDUGAODENGYOUDIAN。
实验步骤
1.PEIZHISHIJI
(1)Lysis Buffer:
Lysis Buffer (5X) YONGSHUIXISHI5BEI,PEIZHICHENGGONGZUOYE;
(2)Luciferase Reaction Reagent:
YONGLuciferase Reaction Buffer(10 ml)RONGJIELuciferase Reaction SubstrateGANFEN(DUANZANLIXINZAIKAIGAI),HUNYUNHOUFENZHUANG、BIGUANGBAOCUN,BIMIANFANFUDONGRONG;
(3)Luciferase Reaction Reagent II:
YONGLuciferase Reaction Buffer II(10 ml)XISHI、HUNHELuciferase Reaction Substrate II(DUANZANLIXINZAIKAIGAI),FENZHUANG、BIGUANGBAOCUN,BIMIANFANFUDONGRONG。
2.LIEJIEXIBAO
XIBAOLIEJIESHIMIDUYINGXIAOYU95%,XIBAOSHENGZHANGGUOMI,YIDAOZHILIEJIEBUCHONGFEN;ZHILISHUNZHUAN、BINGDUGANRANXIBAOHOU,XUPEIYANGYIDUANSHIJIAN(YIBAN48XIAOSHI)SHIYINGGUANGSUMEICHONGFENBIAODA;DIAOJIESHIYANCANSHU,YIMANZUSHANGSHUTIAOJIAN;
QUCHUPEIYANGJI,YONGPBSRUNXIYIBIAN,ZHUYIBUYAOJIANGXIBAOCHUIQI;JIARUSHILIANGDE1X Lysis Buffer,TUIJIANYONGLIANGJIANXIABIAO;
JIANGXIBAOPEIYANGBANZHIYUYAOCHUANGSHANG,SHIWEN、QINGHUANGFUYU15FENZHONG;
JIANGLIEJIEWUZHUANYIDAOXINDEEPGUANZHONG,4℃、12,000rpmLIXIN1FENZHONG,QUSHANGQINGBEIYONG。
3.YINGGUANGJIANCE
QU20μlLIEJIEWU,JIARU100μlPINGHENGZHISHIWENDELuciferase Reaction Reagent,HUNYUNHOUYUMEIBIAOYIZHONGJIANCEfLucXINHAO;
RANHOUJIARU100μlPINGHENGZHISHIWENDELuciferase Reaction Reagent II,HUNYUNHOUYUMEIBIAOYIZHONGJIANCErLucXINHAO。
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