gavin_yan83

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[交流] 【求助/交流】求高手：多肽如何標(biāo)記FITC？

我合成的一個九肽想標(biāo)記FITC，多肽是水溶性的，在DMF中反應(yīng)沒反應(yīng)成功，MS檢測分子量不對。不知有誰做過類似的實驗，想請教一下具體的反應(yīng)條件，最好能給個詳細(xì)的Protocol，先謝謝了！

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1樓 2010-12-21 14:45:16

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zdy1398

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gavin_yan83(金幣+2):謝謝！ 2010-12-25 19:23:46

MS檢測不一定能說明問題，我的多肽連上FITC后MS并沒有檢測出來

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2樓2010-12-24 16:24:43

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zdy1398

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scelab(金幣+2):歡迎多來交流！:tiger06: 2010-12-24 23:20:37
gavin_yan83(金幣+2):謝謝！ 2010-12-25 19:23:56

我的多肽是酯溶的， FITC過量，PH調(diào)到堿性，4℃攪拌過夜··我是這樣做的，僅供參考

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3樓2010-12-24 16:25:53

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xhjggl

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★ ★
scelab(金幣+2):謝謝提供！:tiger06: 2010-12-24 23:20:48

上海生工有進(jìn)口熒光標(biāo)記試劑盒賣。

HOOK™ Dye Labeling Kit (5/6) TAMRA-SE (Rhodamine)
蛋白羅丹明標(biāo)記試劑盒
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℃
編號包裝價格產(chǎn)地
786-058 5 Reactions 2,486 GBios

HOOK™ Dye Labeling Kit (FITC)
蛋白FITC熒光素標(biāo)記試劑盒
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℃
編號包裝價格產(chǎn)地
786-059 5 Reactions 2,486 GBios

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4樓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檢測不一定能說明問題，我的多肽連上FITC后MS并沒有檢測出來

那怎么知道多肽已經(jīng)連上FITC了呢？

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5樓2010-12-25 19:22:54

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zdy1398

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gavin_yan83(金幣+6):謝謝，我已經(jīng)合成好了. 2010-12-29 13:14:00

引用回帖:

Originally posted by gavin_yan83 at 2010-12-25 19:22:54:

那怎么知道多肽已經(jīng)連上FITC了呢？

貌似是這樣的，質(zhì)譜也分很多種，有的能打出來，有的就打不出來，你可以和打質(zhì)譜的老師多交流交流

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6樓2010-12-26 21:48:03

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qq45281873

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小木蟲: 金幣+0.5, 給個紅包，謝謝回帖

請問樓主現(xiàn)在是怎么做的

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7樓2013-04-25 21:13:35

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