H2DCFDA-DCFH-DA-DataSheet-生命科學(xué)試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEH2DCFDACat. No.: HY-D0940CAS No.: 4091-99-0Synonyms: DCFH-DA; 2,7-Dichlorodihydrofluorescein diacetate分式: CHClO分量: 487.29作靶點: Reactive Oxygen Species作通路: Immunology/Inflammation; Metabolic Enzyme/Protease; NF-B儲存式: -20C, protect

2、 from light* In solvent : -80C, 6 months; -20C, 1 month (protect fromlight)溶解性數(shù)據(jù)體外實驗 DMSO : 150 mg/mL (307.82 mM)Ethanol : 14.29 mg/mL (29.33 mM; Need ultrasonic)H2O : 0.1 mg/mL (insoluble)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 2.0522 mL 10.2608 mL

3、 20.5217 mL5 mM 0.4104 mL 2.0522 mL 4.1043 mL10 mM 0.2052 mL 1.0261 mL 2.0522 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 H2DCFDA可滲透細(xì)胞，于檢測細(xì)胞內(nèi)活性氧 (ROS) 的探針。體外研究Analysis of H2DCFDA probe oxidation by means of flow cytometry technique showed that the basal level ofROS in difESC

4、s is about 67 times larger than that in embryonic stem cells (ESCs). Both H2DCFDA and1/2 Master of Small Molecules 您邊的抑制劑師www.MedChemEDCFDA probes are non-fluorescent in their initial form but they undergo multistep conversion inside the cellthat results in the formation of fluorescent product dichl

5、orofluorescein (DCF). The only difference betweenthese two probes is that the conversion of H2DCFDA involves oxidation. Therefore, fluorescence of theH2DCFDA-treated cells depends on the intracellular ROS level, in contrast to fluorescence associated withthe DCFDA probe. Surprisingly, flow cytometry

6、 analysis shows that the difference between fluorescencelevels of ESCs and difESCs loaded with H2DCFDA is close to the difference between the signals fromDCFDA-treated cells. The latter indicates a ROS-independent reason for low fluorescent signal of oxidizedH2DCFDA in ESCs 1.PROTOCOLKinase Assay RO

7、S Measurements 1For the detection of intracellular ROS level ,ROS-sensitive probe H2DCFDA is used. Adherent cells (ESCs,difESCs, eMSCs, HeLa, U118) are incubated with 5 M staining solution in PBS in the dark for 30 min at37C, then harvested with 0.05% trypsin-EDTA solution, suspended in a fresh medi

8、um, and immediatelyanalyzed with flow cytometer. Lymphocytes, both control and PHA-activated, are resuspended in PBS,incubated with 5 M of H2DCFDA in the dark for 30 min at 37C, and immediately analyzed. Along with theH2DCFDA probe, if indicated, ROS-insensitive modification of the fluorescent dye D

9、CFDA is used. Thestaining procedure is the same as for the H2DCFDA 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Cell Rep. 2019 Jul 2;28(1):78-90.e6. EBioMedicine. 2018 Dec. Acta Biomater. 2019 Jun 28. pii: S1742-7061(19)30475-1. Oxid Me

10、d Cell Longev. 2019 Jul. Int J Mol Sci. 2019 Feb 18;20(4). pii: E880.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Lyublinskaya OG, et al. Redox environment in stem and differentiated cells: A quantitative approach. Redox Biol. 2017 Aug;12:758-769.McePdfHeightCaution: Product has not been fully valid