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Cell Meter 細胞內NADH / NADPH熒光成像試劑盒 深紅色熒光

貨號15295存儲條件 在零下15度以下保存, 避免光照
規格100 Tests價格6432
Ex (nm)593Em (nm)655
分子量溶劑
產品詳細介紹


簡要概述

Cell Meter 細胞內NADH / NADPH熒光成像試劑盒 深紅色熒光 是美國AAT Bioquest生產的用于檢測NADH/NADPH的試劑盒,細胞內二氫煙酰胺腺嘌呤二核苷酸NADH及其磷酸酯NADPH的檢測對于疾病診斷和藥物發現是重要的。通常,氧化還原偶聯NAD / NADH和NADP / NADPH在能量代謝,糖酵解,三羧酸循環和線粒體呼吸中起關鍵作用。細胞中NAD(P)H水平的增加與活性氧(ROS)和DNA損傷的異常產生有關。然而,由于缺乏敏感的NAD(P)H探針,在生物系統中檢測細胞內NAD(P)H具有挑戰性。 Cell Meter™細胞內NADH / NADPH熒光成像試劑盒提供了監測遠光譜中活細胞中細胞內NAD(P)H水平的有效方法,并且可以與其他應用如GFP表達細胞或MitoTracker的應用相結合。 JJ1902 NAD(P)H是一種優秀的熒光探針,用于檢測和成像細胞中的NADH / NADPH。該熒光探針結合NADH / NADPH,產生高靈敏度和特異性的強熒光信號。 JJ1902 NAD(P)H傳感器可以很容易地加載到活細胞中,使用Cy5®過濾器可以方便地監測其熒光信號。該試劑盒針對熒光成像和酶標儀應用進行了優化。百螢生物是AAT Bioquest的中國代理商,為您提供最優質的Cell Meter 細胞內NADH / NADPH熒光成像試劑盒。 

 

適用儀器


熒光顯微鏡  
激發: 590nm
發射: 655nm
推薦孔板: 黑色透明
濾波片: Cy5濾波片組


產品說明書

實驗樣品示例

概述

1.在生長培養基中制備細胞

2.將細胞與測試化合物和JJ1902 NAD(P)H傳感器工作溶液在37℃孵育20  -  30分鐘

3.將細胞洗凈并保持在測定緩沖液中

4.在Ex / Em = 590/655 nm(截止= 610 nm)或使用Cy5®過濾器的熒光顯微鏡下監測熒光強度(底部讀取模式)

注意:在開始實驗之前,在室溫下解凍所有試劑盒組分。

 

操作方法

1.制備工作溶液

將10μL JJ1902 NAD(P)H探針儲備溶液(組分A)加入2.5mL測定緩沖液(組分B)中,并充分混合。該JZL1707 NAD(P)H探針工作溶液在室溫下1小時內穩定。

注意:40μL的JZL1707 NAD(P)H傳感器原液足以用于一個板

 

2.實驗步驟

①刺激NADP / NADPH,用10μL10X試驗化合物(96孔板)或5μL5X試驗化合物(384孔板)在無血清培養基或所需緩沖液(如PBS或HHBS)中處理細胞。 對于對照孔(未處理的細胞),加入相應量的培養基或化合物緩沖液。

注意:JJ1902 NAD(P)H探針在血清存在的情況下也是兼容的。探針的條件優化是強烈推薦的細胞系到細胞系。

②在細胞板中加入100μL/孔(96孔板)或25μL/孔(384孔板)的JJ1902 NAD(P)H探針工作溶液。將細胞與測試化合物和JJ1902 NAD(P)H探針工作溶液在37℃共孵育20-30分鐘,避光。

注意:對于NADH / NADPH陽性對照處理:將HeLa細胞與100μMNADH或NADPH在無血清培養基中孵育30分鐘,并與JJ1902 NAD(P)H探針工作溶液在37℃下共培養30分鐘。 詳細信息請參見圖1。

③用所需緩沖液洗滌細胞一次。移除每個孔中的溶液,并添加100μL/孔的測定緩沖液(組分B)用于96孔板或25μL/孔用于384孔板。

④使用底板讀取模式在Ex / Em = 590 / 655nm(截止= 610 nm)下使用酶標儀監測熒光增加,或使用熒光顯微鏡和Cy5®過濾器過濾器獲取圖像。

 

參考文獻

Celastrol attenuates angiotensin II mediated human umbilical vein endothelial cells damage through activation of Nrf2/ERK1/2/Nox2 signal pathway
Authors: Miao Li, Xin Liu, Yongpeng He, Qingyin Zheng, Min Wang, Yu Wu, Yuanpeng Zhang, Chaoyun Wang
Journal: European Journal of Pharmacology (2017): 124--133

Cytosolic Redox Status of Wine Yeast (Saccharomyces Cerevisiae) under Hyperosmotic Stress during Icewine Fermentation
Authors: Fei Yang, Caitlin Heit, Debra L Inglis
Journal: Fermentation (2017): 61

Epigenetic regulation of Runx2 transcription and osteoblast differentiation by nicotinamide phosphoribosyltransferase
Authors: Min Ling, Peixin Huang, Shamima Islam, Daniel P Heruth, Xuanan Li, Li Qin Zhang, Ding-You Li, Zhaohui Hu, Shui Qing Ye
Journal: Cell & Bioscience (2017): 27

MCU-dependent mitochondrial Ca2+ inhibits NAD+/SIRT3/SOD2 pathway to promote ROS production and metastasis of HCC cells
Authors: T Ren, H Zhang, J Wang, J Zhu, M Jin, Y Wu, X Guo, L Ji, Q Huang, H Yang
Journal: Oncogene (2017)

Metabolic and molecular insights into an essential role of nicotinamide phosphoribosyltransferase
Authors: Li Q Zhang, Leon Van Haandel, Min Xiong, Peixin Huang, Daniel P Heruth, Charlie Bi, Roger Gaedigk, Xun Jiang, Ding-You Li, Gerald Wyckoff
Journal: Cell Death & Disease (2017): e2705

Pyrroloquinoline Quinone, a Redox-active o-Quinone, Stimulates Mitochondrial Biogenesis by Activating SIRT1/PGC-1α Signaling Pathway
Authors: Kazuhiro Saihara, Ryosuke Kamikubo, Kazuto Ikemoto, Koji Uchida, Mitsugu Akagawa
Journal: Biochemistry (2017)

Resveratrol attenuates excessive ethanol exposure induced insulin resistance in rats via improving NAD+/NADH ratio
Authors: Gang Luo, Bingqing Huang, Xiang Qiu, Lin Xiao, Ning Wang, Qin Gao, Wei Yang, Liping Hao
Journal: Molecular Nutrition & Food Research (2017)

A Snapshot of the Plant Glycated Proteome STRUCTURAL, FUNCTIONAL, AND MECHANISTIC ASPECTS
Authors: Tatiana Bilova, Elena Lukasheva, Dominic Brauch, Uta Greifenhagen, Gagan Paudel, Elena Tarakhovskaya, Nadezhda Frolova, Juliane Mittasch, Gerd Ulrich Balcke, Alain Tissier
Journal: Journal of Biological Chemistry (2016): 7621--7636

AMPK activation protects cells from oxidative stress-induced senescence via autophagic flux restoration and intracellular NAD+ elevation
Authors: Xiaojuan Han, Haoran Tai, Xiaobo Wang, Zhe Wang, Jiao Zhou, Xiawei Wei, Yi Ding, Hui Gong, Chunfen Mo, Jie Zhang
Journal: Aging cell (2016): 416--427

Cell-Line Selectivity Improves the Predictive Power of Pharmacogenomic Analyses and Helps Identify NADPH as Biomarker for Ferroptosis Sensitivity
Authors: Kenichi Shimada, Miki Hayano, Nen C Pagano, Brent R Stockwell
Journal: Cell chemical biology (2016): 225--235

 

相關產品

產品名稱 貨號
Cell Meter 胞內NADH / NADPH熒光成像分析試劑盒 Cat#15290


說明書
Cell Meter 細胞內NADH / NADPH熒光成像試劑盒 深紅色熒光 .pdf


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