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HOME > Products & Services > ROSFluor™ Series > [Measurement of reactive oxgen species]Hydroxyphenyl Fluorescein (HPF) / [Measurement of reactive oxgen species]Aminophenyl Fluorescein (APF)

[Measurement of reactive oxgen species]Hydroxyphenyl Fluorescein (HPF) / [Measurement of reactive oxgen species]Aminophenyl Fluorescein (APF)

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.

Hydroxyphenyl Fluorescein(HPF) / Aminophenyl Fluorescein(APF)

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.

Hydroxyphenyl Fluorescein(HPF) / Aminophenyl Fluorescein(APF)

 

Hydroxyphenyl Fluorescein(HPF) / Aminophenyl Fluorescein(APF)   Hydroxyphenyl Fluorescein(HPF) / Aminophenyl Fluorescein(APF)

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 Download
545KB
Download
127KB
SK3002-01 Aminophenyl Fluorescein (APF) 1mg $459 Download
132KB