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小木虫论坛-学术科研互动平台 » 生物医药区 » 分子生物 » 综合其他 » 【求助/交流】求高手：多肽如何标记FITC？
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gavin_yan83

木虫 (正式写手)

[交流] 【求助/交流】求高手：多肽如何标记FITC？

我合成的一个九肽想标记FITC，多肽是水溶性的，在DMF中反应没反应成功，MS检测分子量不对。不知有谁做过类似的实验，想请教一下具体的反应条件，最好能给个详细的Protocol，先谢谢了！
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1楼 2010-12-21 14:45:16
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zdy1398

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scelab(金币+2):欢迎多来交流！:tiger06: 2010-12-24 23:20:37
gavin_yan83(金币+2):谢谢！ 2010-12-25 19:23:56
我的多肽是酯溶的， FITC过量，PH调到碱性，4℃搅拌过夜··我是这样做的，仅供参考
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3楼2010-12-24 16:25:53
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zdy1398

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gavin_yan83(金币+2):谢谢！ 2010-12-25 19:23:46
MS检测不一定能说明问题，我的多肽连上FITC后MS并没有检测出来
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2楼2010-12-24 16:24:43
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xhjggl

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scelab(金币+2):谢谢提供！:tiger06: 2010-12-24 23:20:48
上海生工有进口荧光标记试剂盒卖。

HOOK™ Dye Labeling Kit (5/6) TAMRA-SE (Rhodamine)
蛋白罗丹明标记试剂盒       
Description:
(5/6) TAMRA-SE (5-(and-6)-Carboxytetramethylrhodamine succinimidyl ester, mixed isomers) is based on tetramethylrhodamine, one of the most common fluorophores used in the labeling of peptides, proteins, nucleic acids and nucleotides.(5/6) TAMRA absorbs green visible light at 546nm and emits an orange-red visible light at a maximum emission of 575nm.The NHS ester group provides the simplest and most commonly used group for labeling proteins. The succinimidyl ester group reacts with primary amines in lysine side chains and N-terminal amines forming a stable, covalent amide bond.This kit utilizes Tube-O-REACTOR™, which allows for the reactions to be performed in a single tube, with no loss of essential reagents and minimum hands-on time and effort. The (5/6) TAMRA-SE labeling reagent is also available in our single use OneQuant™ format.
Features:
Dye preweighed and supplied in single use  vials (Also available separately);Suitable for most proteins;Utilizes  single Tube Reactor for minimum hands on time and no protein loss.application for Labeling of proteins, peptides and nucleic acids with a red fluorescent dye.;Suitable for antibody labeling.Store: -15～-20℃
编号        包装        价格        产地
786-058        5 Reactions        2,486        GBios

HOOK™ Dye Labeling Kit (FITC)
蛋白FITC荧光素标记试剂盒       
Description:
FITC (fluorescein isothiocyanate) is a commonly used fluorescent label for proteins, as it contains the groups required for conjugating to amino, sulfhydryl, imidazoyl, tyrosyl or carbonyl groups of proteins. FITC has a molecular weight of 389, and excitation and emission wavelengths of 494nm and 520nm, respectively, therefore emitting green visible light.This kit utilizes Tube-O-Reactor™, which allows for the reactions to be performed in a single tube, with no loss of essential reagents and minimum hands-on time and effort.
Features:
Dye preweighed and supplied in single use  vials (Also available separately);Suitable for most proteins;Utilizes single Tube Reactor for minimum hands on time.application for Labeling of a green fluorescent dye to proteins and peptides. And Suitable for antibody labeling. Store: -15～-20℃
编号        包装        价格        产地
786-059        5 Reactions        2,486        GBios
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4楼2010-12-24 21:48:43
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gavin_yan83

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引用回帖:
Originally posted by zdy1398 at 2010-12-24 16:24:43:
MS检测不一定能说明问题，我的多肽连上FITC后MS并没有检测出来

那怎么知道多肽已经连上FITC了呢？
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5楼2010-12-25 19:22:54
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