Detection of Glycoproteins on Blot

Source:ContributedbySharadPurohit,Paller"sLab

Reagents SodiumacetateBuffer(200mM,pH5.5):Preparea200mMsolutionofsodiumacetatebydisolving12.0gamofsodiumacetatetrihydratein440mlofdw.Preparea200mMsolutionofaceticacidbydissolving690ulofglacialaceticacidto60mlDw.MixthetwosolutionandcheckpH PBSpH7.2:dissolve575mgdisodiumphosphate,100mgofsodiumdihydrogenphosphateand800mgsodiumchloridein500mlDw. Sodiumacetatebuffer/5mMEDTA:dissolve1.14gofEDTA(Tetrasodiumsalt)in100mlof200mMsodiumacetatebufferanddiluteto200mlwithDw 10mMsodiummetaperiodatesolutionin100mMsodiumacetate/5mMEDTA BiotinHydrazide:Prepareastocksolutionof1mg/mlinDMF.Workingsolutionis100ug/mlin100mMSodiumacetatebuffercontaining5mMEDTApH5.5.Preparefresheverytime TBS-T BlockingSolution:;5%gelatininTBS-T. Colordevelopmentreagents Procedure WashtheblotextensivelywithPBStoremovetracesoftris(astrisinterfereswithbiotinylationreaction). Immersetheblotin10mlof10mMsodiumperiodatesolutionandincubateindarkfor25minutes. Washmembrane3timeswithPBSfor10minuteseachwasatroomtemperaturewithgentleagitation. Preparetheworkingbiotinylationsolutionjustbeforeuseandincubatethemembraneintheisolutionfor90minutesatroomtemperaturewithgentleagitation Washthemembrane5timeswithTBS-T(TBS-Twillnolongerintereferewiththeprocessfromthisstageonwards)for20minuteswithgentleagitation Blockthemembraneinblockingsolutionfor1hourat37degC. WashmembranewithTBS-TandaddthesecondaryantibodyconjugatedwithAlkalinePhosphataseorHRPfor1hour. WashthemembranewithTBS-T. Developblotwiththedevlopmentreagentofthechoicetillyougetadesiredcontrast. Precautions Tris,2-mercaptoethanol,ammoniumions,DTT,monosaccharides,sodiumborohydridetranstionmetalsandureaaretheinterfereingcompunds.Henceavoidtheirusebeforethebiotinylationreaction. 您的称呼： 单位名称： 您的邮箱： 您的电话： 品牌与型号： 其他要求： 新闻动态 行业前沿 技术文章 最新产品 New complete structure of Hafnia alvei clinical isolate strain PCM... 2021-03-26 Polish Collection of Microorganisms一级代理 pcm中国 2021-04-08 进口Polish Collection of Microorganisms品牌清关代理 pcm产品促销 2018-01-11 Polish Collection of Microorganisms中国采购 Polish Collection of 2017-11-23 Core Oligosaccharide of Plesiomonas shigelloides PCM 2231... 2021-09-13 Polish Collection of Microorganisms代理，产品目录 pcm试剂产品信息 2018-05-29 Polish Collection of Microorganisms试剂产品信息 pcm报价单 2017-01-15 pcm代理 pcm授权 2017-10-10 Polish Collection of Microorganisms公司,Polish Collection of Micro... 2019-07-07 Polish Journal of Microbiology 期刊投稿经验分享,Polish... 2018-03-27 MPB:湖南农大尹杰组猪粪便来源乳酸菌分离技术_刘永鑫的博客—... 2017-10-27 IDA: 波兰 (Polish Collection of Microorganisms (PCM)) 2018-11-09 CP到底什么意思? 2019-07-08 Enrichment, Isolation and Susceptibility Profile of the... 2019-08-14 Core Oligosaccharide of Plesiomonas shigelloides PCM 2231... 2020-07-09 www.csdn.net 2019-06-13 Core Oligosaccharide of Plesiomonas shigelloides PCM 2231... 2021-09-12 【求助】加酶量与底物浓度比值的计算方法 食品 学术科研... 2021-09-15 各国菌种保藏机构_我们都爱微生物 2021-09-23 决定抗原免疫原性的因素有哪些?怎样才能获得高效价的抗体? 上学... 2021-09-21 《德语助手》德语在线词典_德语翻译_德语动词变位_德语 ... 2021-09-20 G01.595.210 [Categoria DeCS] 2021-09-14 收藏帖!国内外菌种保藏机构_网易订阅 2021-09-16 Core Oligosaccharide of Plesiomonas shigelloides PCM 2231... 2021-09-22 波兰微生物/肺炎克雷伯菌/PCM 2713 2026-07-06 波兰微生物/肺炎克雷伯菌/PCM 2070 2026-07-06 波兰微生物/肺炎克雷伯菌/PCM 2068 2026-07-06 波兰微生物/肺炎克雷伯菌鼻硬化症/PCM 2067 2026-07-06 波兰微生物/肺炎克雷伯菌/PCM 2064 2026-07-06 波兰微生物采集/肺炎克雷伯菌/PCM 2069 2026-07-06 Copyright@苏州蚂蚁淘生物科技有限公司 2003-2021  188进口试剂采购网版权所有     苏ICP备17049038号-9 友情链接：  其他 188进口试剂采购网 www.188bio.cn -中国试剂网,试剂网,化学试剂网,国药试剂,抗体公司,试剂公司,试剂盒公司,苏州试剂公司,北京化学试剂公司,天津化学试剂,试剂商城,试剂代理,流式抗体 细胞库查询 sitemap