目錄号 | MX5203-1MG | 售價 | 339.00元 | ||||||||||||||||||||
規格 | 1mg | 運輸溫度 | 冰袋運輸 | ||||||||||||||||||||
其他名稱 | N-Acetyl-3,7-dihydroxyphenoxazine N-乙酰基-3,7-二羟基吩嗪;ADHP;Amplex® Red Reagent;A 6550;Ampliflu™ Red 熒光(guāng)紅(hóng)染 | 保存溫度 | -20℃避光(guāng)幹燥保存 | ||||||||||||||||||||
CAS号 | 119171-73-2 | 有效期 | 2年(nián) | ||||||||||||||||||||
應用 | 過氧化氫探針 | 訂購數量 |
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産品簡介: 10-Acetyl-3,7-dihydroxyphenoxazine (ADHP) 過氧化氫探針 (Amplex® Red H2O2 probe 過氧化氫探針) 搜索關鍵詞: 10-Acetyl-3,7-dihydroxyphenoxazine (ADHP);Amplex®Red; Resorufin 試鹵靈;過氧化氫探針;Ampliflu™ Red;CAS NO:119171-73-2;
基本介紹: 10-Acetyl-3,7-dihydroxyphenoxazine (ADHP),中文名10-乙酰基-3,7-二羟基吩嗪,以商标名Amplex®Red在市(shì)場上(shàng)銷售。ADHP是一(yī)種高(gāo)度靈敏和穩定的過氧化物(wù)酶底物(wù),選擇性檢測過氧化氫(H2O2)。存在辣根過氧化物(wù)酶(HRP)的體系中,這一(yī)基本無色和非熒光(guāng)的ADHP與H2O2以1:1的化學定量比反應,生(shēng)成強紅(hóng)色熒光(guāng)的試鹵靈(Resorufin)(Ex/Em= 571/585nm),可通(tōng)過熒光(guāng)光(guāng)度計和酶标儀分析結果。許多(duō)酶反應能(néng)産生(shēng)H2O2,因此,ADHP能(néng)夠用在偶聯酶促反應中檢測許多(duō)不同酶的生(shēng)物(wù)活性。
工(gōng)作原理:
Fig 1. 使用ADHP進行偶聯酶促反應的工(gōng)作原理。葡萄糖氧化酶催化葡萄糖氧化生(shēng)成過氧化氫(H2O2),通(tōng)過辣根過氧化物(wù)酶(HRP)催化ADHP反應生(shēng)成強熒光(guāng)試鹵靈(Resorufin)。
産品應用: 10-Acetyl-3,7-dihydroxyphenoxazine (ADHP)是目前位置最靈敏和穩定的熒光(guāng)探針用來檢測HRP和H2O2。 A) ADHP普遍用來檢測許多(duō)免疫實驗中的HRP; B) ADHP用來檢測微量的H2O2;
Zhou, M.等研究表明ADHP(Amplex®Red)檢測過氧化物(wù)酶活性,最少能(néng)檢測1×10−5U/mL HRP;或H2O2,最少能(néng)檢測50nM H2O2。【文獻來源:Zhou M et al. A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide: applications in detecting the activity of phagocyte NADPH oxidase and other oxidases. Anal Biochem. 1997 Nov 15;253(2):162-8.】
基本特性: 1) CAS NO.:119171-73-2 2) 化學名:1-(3,7-dihydroxy-10H-phenoxazin-10-yl)-ethanone; 10H-Phenoxazine-3,7-diol, 10-acetyl-; 3) 同義名:N-Acetyl-3,7-dihydroxyphenoxazine N-乙酰基-3,7-二羟基吩嗪;ADHP;Amplex®Red Reagent;A 6550;Ampliflu™ Red 熒光(guāng)紅(hóng)染料; 4) 分子式:C14H11NO4 5) 分子量:257.25 6) 純度:≥98% 7) 摩爾消光(guāng)系數(ε):54,000 (Resorufin) 8) Ex/Em:571/585nm (pH 9.0, Resorufin) 9) 溶解性:溶于DMSO、DMF 10) 化學結構式:
ADHP的應用實例: 文獻來源:Summers FA et al. Photooxidation of Amplex Red to resorufin: implications of exposing the Amplex Red assay to light. Methods Enzymol. 2013;526:1-17.
實驗1)Amplex Red Assay Reactions containing 50 µM Amplex Red reagent, 0.1 U/mL HRP, H2O2 and 0.5 U/mL catalase in 50 mM sodium phosphate buffer, pH 7.4, as indicated in the figure legends, were prepared in a darkroom under a red light. White ELISA plates containing the samples (100 µL/well) in triplicate were either kept in the dark for 30 and 60 min or under ambient laboratory light. The fluorescence (excitation 535 nm, emission 595 nm) was measured on a GENios platereader (Tecan, Durham, NC, USA).
實驗2)Cell Culture Assay HaCaT keratinocytes were incubated with 50 µM Amplex Red in PBS for 1 h at 37°C in the dark, then cells were irradiated using UVA (315–400 nm) from four fluorescent PUVA lamps (Houvalite F20T12BL-HO; National Biological Co., Twinsburg, OH) for 15 minutes (3.75 J/cm2) and 30 minutes (7.5 J/cm2). Control cells were treated with the same conditions. After exposure, the fluorescence (excitation 535 nm, emission 595 nm) was recorded using a plate reader.
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注意事(shì)項 1) ADHP的使用比較方便,可配制10-20mM DMSO儲存液,之後用反應緩沖液比如PBS,Tris buffer稀釋到(dào)50-200µM工(gōng)作濃度用于檢測。最佳的工(gōng)作濃度需根據自(zì)身實驗需求進行優化。 2) Amplex®Red是Life Technologies公司的注冊商标名。 3) 為(wèi)了您的安全和健康,請穿實驗服并戴一(yī)次性手套操作。
— —Written/Edited by V. Shallan【版權歸MKBio懋康所有】
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