Dihydrorhodamine 123 (C008B-266956)

Dihydrorhodamine 123 (DHR123) is a non-charged and non-fluorescent reactive oxygen species (ROS) indicator that can diffuse across cell membranes. After entering cells, in the presence of oxygen free radicals, it is oxidized to rhodamine 123 cation in mitochondria and emits green fluorescence (Ex/Em= 507/529 nm). DHR123 is commonly used to detect reactive oxygen intermediates produced by endothelial cells, eosinophils, and reactive microglia.Staining Procedure(1) Dilute the 5 mM DHR123 stock solution 50-fold with culture medium to obtain a 100 µM DHR123 working solution.(2) Prepare two batches of HL60 cells at a cell density of ~6×10⁶ cells/mL, using Krebs-Ringer buffer as the buffer system.(3) Add DHR123 working solution at 1/10 the volume of HL60 cell suspension to both batches, yielding a final DHR123 concentration of 10 µM, and incubate at 37 ℃ for 30 minutes.(4) To one batch of HL60 cells after incubation with DHR123, add the oxygen free radical inducer phorbol 12-myristate 13-acetate (PMA) to a final concentration of 50 µM.(5) To the other batch of HL60 cells, add SNARF-1 AM acetate (SNARF-AM) probe to a final concentration of 5 µM, incubate at 37 ℃ for 10 min, centrifuge, and wash the cells with culture medium.(6) Observe the cells under a fluorescence microscope equipped with a fluorescein filter set.Precautions(1) Prepare solutions fresh before use.(2) For your safety and health, please wear a lab coat and disposable gloves.ReferenceLydia M. Henderson et al. Dihydrorhodamine 123: a fluorescent probe for superoxide generation. Eur. J. Biochem. 217, 973-980.

Specifications & Purity: Dihydrorhodamine 123 (DHR123) is a non-charged and non-fluorescent reactive oxygen species (ROS) indicator that can diffuse across cell membranes. After entering cells, in the presence of oxygen free radicals, it is oxidized to rhodamine 123 cation in mitochondria and emits green fluorescence (Ex/Em= 507/529 nm). DHR123 is commonly used to detect reactive oxygen intermediates produced by endothelial cells, eosinophils, and reactive microglia.Staining Procedure(1) Dilute the 5 mM DHR123 stock solution 50-fold with culture medium to obtain a 100 µM DHR123 working solution.(2) Prepare two batches of HL60 cells at a cell density of ~6×10⁶ cells/mL, using Krebs-Ringer buffer as the buffer system.(3) Add DHR123 working solution at 1/10 the volume of HL60 cell suspension to both batches, yielding a final DHR123 concentration of 10 µM, and incubate at 37 ℃ for 30 minutes.(4) To one batch of HL60 cells after incubation with DHR123, add the oxygen free radical inducer phorbol 12-myristate 13-acetate (PMA) to a final concentration of 50 µM.(5) To the other batch of HL60 cells, add SNARF-1 AM acetate (SNARF-AM) probe to a final concentration of 5 µM, incubate at 37 ℃ for 10 min, centrifuge, and wash the cells with culture medium.(6) Observe the cells under a fluorescence microscope equipped with a fluorescein filter set.Precautions(1) Prepare solutions fresh before use.(2) For your safety and health, please wear a lab coat and disposable gloves.ReferenceLydia M. Henderson et al. Dihydrorhodamine 123: a fluorescent probe for superoxide generation. Eur. J. Biochem. 217, 973-980.

Grade: BioReagent;Biological Stain;for Fluorescence analysis;for Microscopy

Molecular Formula: C21H18N2O3

Molecular Weight: 346.38

EC Number: 886-538-4

PubChem CID: 105032

Isomeric SMILES: COC(=O)C1=CC=CC=C1C2C3=C(C=C(C=C3)N)OC4=C2C=CC(=C4)N

Storage Temperature: Protected from light;Store at -20°C

Shipped in: Ice chest + Ice pads

UPC:

51373462

Weight:

1.09 Ounces

HazmatClass:

Yes

Quantity:

1

Unit of Measurement:

EA

WeightUOM:

LB

MPN:

D1520930-50μl

Country of Origin:

China

CAS:

109244-58-8

Product Size:

50μl

Harmonized Tariff Code:

2931.90.90