Features

• HFP and AFP detect selectively highly reactive oxygen species (hROS) such as hydroxyl radical (・OH) and peroxynitrite (ONOO－), whereas they do not react with other reactive oxygen species (O－・₂ , H₂O₂, ¹O₂, NO, etc.).
• Quantitative analysis of H₂O₂ is available by adding HRP.
• Detecting specifically hypochlorite (OCl－) by the combination use of HPF and APF.
• HPF and APF are not autoxidized at all, which enables us to easily get more reliable data than by other reagents.
• Live fluorescence imaging of cells is available.

Principle of the measurement

HPF and APF are almost non-fluorescent in neutral solution. When they react with highly reactive oxygen species (hROS), they generate fluorescein that has high fluorescence intensity  (excitation wave length: 490 nm, fluorescent wave length: 515 nm), and increase in the intensity of the fluorescence is observed.

Fluorescent probes for detecting reactive oxygen species (ROS)

• Each fluorescent reagent (final concentration: 10 mM, added 0.1 % DMF as co-solvent) was reacted with various reactive oxygen species in phosphate buffer (0.1 M, pH 7.4).
• Fluorescent intensity of HPF, APF and DCFH was measured at 515, 515, 520nm by the excitation of 490, 490, 500 nm, respectively.

Preparation of the reagent

1mg of HPF / APF is dissolve in N,N -dimethylformamide（DMF） 0.47 mL. Their densities are 5 mmol/L, each. Dilute the reagent 500～5000 times （10～1μmol/L） by Phosphate buffer (0.1 mol/L, pH 7.4) before use. Adding the diluted reagent to the sample and incubate, then measure the fluorescence of wave length: 515 nm with excitation of wave length: 490 nm.

Application Note

Reference

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price

Code no.	Product	size	Price	Protocol	MSDS
SK3001-01	Hydroxyphenyl Fluorescein (HPF)	1mg	$459	545KB	127KB
SK3002-01	Aminophenyl Fluorescein (APF)	1mg	$459		132KB

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