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Fluoro MPOHOCL™
Fluorescent Myeloperoxidase  Chlorination and peroxidation Detection Kit
New! 
Key Benefits:
  • Can monitor multiple time points to follow kinetics.
  • One-step, no wash assay.
  • Adaptable for High Throughput format
  • Highly Sensitive
  • Applications - Fluorescence Plate Reader
Assay Principle

Myeloperoxidase (MPO) is a highly cationic glycosolated hemoprotein that has a molecular weight of 144kD. The hemoprotein consists of two dimers linked via a disulfide bridge. Each dimmer is composed of a heavy (53kD) and light (15kD) subunit. Each heavy chain contains an independently acting protoporphyrin group containing a central iron (1-5). MPO is present in the azurophilic granules of polymorphonuclear leukocytes (PMNs) and is unique to neutrophils and monocytes. However, monocytes contain only one third of the MPO found in PMN’s. MPO utilizes H202 produced by the neutrophils to oxidize a varity of aromatic compounds to give substrate radicals for bactericidal activity (4 review). This enzyme is unique however in that it can oxidize chloride ions to produce a strong nonradical oxidant,HOCl. HOCl is the most powerful bactericidal produced by neutrophils (4 review). Excessive production of these radicals can cause oxidative stress leading to oxidative tissue injury.

Chlorination Reaction:

H2O2 + MPO + Cl- ----> HOCl + APF (non fluorescent) ----> Fluorescent Dye
                                          
Excitation:488nm; Emission: 515-530nm

Peroxidation Reaction:

H2O2 + Detection reagent (non-fluorescent)+ MPO ----->   fluorescent analog

 Excitation 530-571nm Emission  590-600nm

Applications
  • Detection of MPO chlorination activity in neutrophils and macrophages.
  • Detection of PMN infiltration in tissue samples (inflammation and innate host defense mechanisms).  
  • Acute and chronic inflammatory disorders due to chlorination tissue damage.  
  • MPO chlorination activity in acute and chronic manifestations of cardiovascular disease.
Kit contents (for 500 assays of Myeloperoxidase detection and 300 assays of Chlorination detection)
  1. 10X Assay Buffer: 60ml 
  2. Detection reagent: One vial for 500 assays.
  3. Hydrogen Peroxide: 1000µL of a stabilized 3% solution.
  4. Myeloperoxidase: 1 vial at 30 units/ml
  5. 1 vial of APF

 

Peroxidase and Chlorination Detection
Catalog 
 
Size
 
Price (Eur)
MPOHOCL 100-3
 
500 + 300 Tests €590