Dichlorofluorescein diacetate

Soybean (Glycine max) direct apphcation of pressure to slide-moimted cells osmotic stress pyranine fluorescence 2, 7 -dichlorofluorescein diacetate [183]... 

Material required Stock solution of 25 mM H2DCFDA, carboxy-2, 7 -dichlorofluorescein diacetate (Sigma Co) dissolved in dimethyl sulphoxide (DMSO). Confocal Microscope BIORAD 1024, Laser KrAr with inverted microscope Nikon TMD 300. 

EDTA, Ethylenediaminetetraacetic acid TAS, total antioxidant status TRAP, total peroxyl-radical-trapping antioxidant capability of plasma ABAP, 2,2 -azobis(2-amidopropane) FRAP, ferric-reducing activity of plasma H2-DCF-DA, 2, 7 -dichlorofluorescein diacetate ORAC, oxygen-radical-absorbing capacity ABTS, 2,2 -azinobis(3-ethylbenzthiazoline-6-sulfonic acid). 

We have previously shown that 4-hydroxynonenal (4-HNE) induced oxidative stress and apoptotic cell death in Swiss 3T3 fibroblasts. Indeed, in vitro treatment of 3T3 fibroblasts with 4-HNE induced a condition of oxidative stress as monitored by the oxidation of dichlorofluorescein diacetate. Further, 4-HNE-treated fibroblasts eventually underwent apoptotic death as determined by differential staining and intemucleosomal DNA fragmentation. These observations are consistent with a potential role of lipid peroxidation-derived products in programmed cell death. " ... 

Intracellular ROS was measured using 2 ,7 -dichlorofluorescein diacetate (DCFH-DA) as described previously (20). MNCs were pretreated with 100 juM DCFH-DA for 1 h, and treated with Trp-P-1 for another 1 h. The cells were twice washed with PBS, and formed 2 ,7 -dichlorofluorescein was spectrofluorometrically measured at excitation and emission wavelengths of 495 and 530 nm, respectively. 

Myhre, O. Andersen, J. M. Aarnes, H. Fonnum, F. Evaluation of the probes 2, 7 -dichlorofluorescein diacetate, luminol, and lucigenin as indicators of reactive species formation. Biochem. Pharmacol. 2003, 65, 1575-1582. 

At the end of treatment, HL-60 cells were stained with 40 nM 3,3 -dihexyloxacarbocyanine (DiOC6 (3), Molecular Probes, Eugene, OR) for 15 min at 37 C. Mitochondrial transmembrane potential was measured by flow cytometry. At least 10,000 cells were collected in each sample. Generation of ROS was monitored with 20 pM 2, 7 -dichlorofluorescein diacetate (DCFH-DA) (Molecular Probes, Eugene, OR) staining for 1 h at 37 C and assayed by flow cytometry. 

Other Names 5-Carboxy-2, 7 -dichlorofluorescein diacetate Carboxy-DCFDA 5-CDCF DA Merck Index Number Not listed Chemical/Dye Class Xanthene Molecular Formula C25H14CI2O9... 

Percy, M. E. Zhao, Y. Lukiw, W J. Metal-sulfate induced generation of ROS in human brain cells detection using an isomeric mixture of 5-and 6-carboxy-2, 7 -dichlorofluorescein diacetate... 

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