Lumiprobe free radical analysis h2dcfda instructions

AI Meijie  Lumiprobe H2DCFDA（2',7'- Dichlorodihydrofluorescein diacetate ） It is a common reagent , It is used to study the production of reactive oxygen species in living cells .

H2DCFDA  It is a non fluorescent fluorescein derivative （ Its reduced acetylated form ）. The reagent is only broken down in acetyl group and the reagent is oxidized in cells and transformed into  2',7'- Dichlorofluorescein starts to emit fluorescence . This is a bright green fluorescent dye （ Maximum absorption  511 nm, Maximum fluorescence  533 nm）. The reagent can be used for determination in living cells , Incompatible with sample fixation .

H2DCFDA  The acetyl group in increases its lipophilicity and improves its cell membrane permeability . Once inside the cell , The dye will be deacetylated by cellular esterase , Thus, it is charged and better fixed in the cell . Oxidation in the form of reactive oxygen species leads to fluorescent products （2',7'- Dichlorofluorescein ） The formation of , And various methods can be used to detect , For example, use flow cytometry 、 Plate reader or fluorescence microscope .

Use emcee  Lumiprobe H2DCFDA The advice of ：

Use the newly prepared reagent solution （ Working solution is not suitable for long-term storage , Because the reagent will gradually oxidize ）.

Specific cell lines and assay conditions , Select the best reagent working concentration and incubation time required for reagent deacetylation and oxidation . If there is no recommended regimen for a specific cell line , From  1  To  10 μM  The concentration of starts and incubates  30  minute .

Do not incubate dyes with cells in the presence of serum , Because it contains cutting  H2DCFDA  Enzymes .

AI Meijie  Lumiprobe H2DCFDA chemical property ：

form ：  Colorless solid

Molecular weight ： 487.29

CAS Number ： 4091-99-0

Molecular formula ： C 24 H 16 Cl 2 O 7

IUPAC  name ：2-(3,6- Diacetoxy -2,7- Dichloride -9H-xanthen-9-yl) Benzoic acid

solubility ：  good  DMSO、 Ethanol and  DMF

The quality control ：NMR 1 H, HPLC-MS (95%)

Storage conditions ：-20°C  After delivery under dark conditions  12  Months . transport ： At room temperature for  3  Zhou . Moisture-proof . Avoid storage without light and unnecessary freezing and thawing cycle of stock solution . If the reagent is used many times , It is strongly recommended to store it in a dry argon or nitrogen atmosphere .

Spectral characteristics

Maximum excitation / absorption ,nm： 511

ε, L⋅mol -1 ⋅cm -1： 118626

Maximum emission , nanometer ： 533

Fluorescence quantum yield ： 0.76

CF 260： 0.17

CF 280： 0.14

AI Meijie  Lumiprobe H2DCFDA Related research ：

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