Myeloperoxidase Detection 

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hROS Detection Kit
Key Benefits:
  • Quenched Cell permeable dye
  • Can be used with Cell Lysates, Tissue Homogenates
  • One Step, No wash Homogenous assay.
  • Adaptable to High throughput assay platforms
  • Can monitor multiple time points to follow real time kinetics
  • Non-destructive cell based assay allows monitoring of additional parameters
  • Applications  - Fluorescence Plate Reader / Fluorescent Microscope / Flow Cytometry
Introduction:
A new novel probe, Aminophenyl fluorescein (APF), developed by Tetsuo Nagano et. al. (1), is a general selective indcator for the detection of highly Reactive Oxygen Species (hROS). The probe has little reactivity towards other ROS such as: singlet oxygen (O21), superoxide (O2– •), hydrogen peroxide (H2O2), nitric oxide (NO•), and alkyl peroxide (RO2•) (see table below)1. APF is a cell permeable indicator that can be used to detect Hydroxyl Radical (.OH), Peroxynitrite: (ONOO- ) and hypochlorite (-OCl) production in cells.  
 
Several fluorescent probes such as 2’,7’-dichlorodihydrofluorescein (DCFH) and di-hydrorhodamine123, have been developed to detect Reactive Oxygen Species (ROS). As Hempel and co-workers (2) have indicated, DCFH and di-hydrorhodamine123 can react with various ROS and oxidizing species such as hydrogen peroxide, hypochlorite, superoxide, nitric oxide, ferrous ion, and others. These probes should be considered as detecting a broad range of ROS (2).
Assay Principle:
Reactivity:
Reactivity Profile of APF:
ROS (RFU)
 		 APF (RFU)
Ex:499 Em:515
 		 DCFH-DA (RFU)
Ex:500 Em:520
 
Hydroxyl Radical: •OH
 		 1200 7400
Peroxynitrite: ONOO- 560
 		 6600
Hypochlorite: -OCl 3600 86
Oxygen Radical: 1O2 9 26
Superoxide: O2-• 6 67
Hydrogen Peroxide : H202 <1 190
Nitric Oxide: NO <1 150
Alkylperoxyl Radical: ROO• 2 710
Autoxidation <1 2000
The following kits are available:
Catalog
 		 Contents Size Price
FLAPF 100-2
 		 APF 1 vial 150 Tests €195
Dilution of APF
 		 Volume of test Numer of tests
1:500 (10mM final)
 		 0.2 mL 147
1: 1000 (5mM final) 0.2mL 294