Eosinophil Peroxidase Detection

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Fluoro EPO
Fluorescent Eosinophil Peroxidase Detection Kit

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) or Absorbance

Assay Principle

Eosinophil peroxidase (EPO) is the most abundant enzyme found in eosinophils. It is the major cytotoxic agent released by activated eosinophils and uses hydrogen peroxide to generate reactive oxidants from halides and pseudo halide thiocyanate [1,2]. Eosinophils peroxidase has been shown to have antimycobacterial activity [3], however it is also implicated in tissue damage that occurs in asthma and other diseases [4,5]. Currently, the function of eosinophil involvement in the immune response is being redefined. Once considered a cell involved in host protection of parasitic infection, eosinophils multiple functions as leukocytes involved in the initiation and propagation of diverse inflammatory responses are being investigated. Eosinophils are further involved as modulators of innate and adaptive immunity [6].

H₂O₂ + Detection reagent (non-fluorescent)+ EPO
---------> fluorescent analog
Excitation:530-571nm; Emission: 590-600nm

Applications

Detection of EPO activity in isolated eosinophils.
Functional studies of eosinophil degranulation.
In-vitro eosinophil chemotaxis studies.

Kit contents (for 100 assays)

5X Reaction Buffer: 60ml ..........................................Part 3011
Detection reagent: One vial for 100 assays....................Part 4007
Hydrogen Peroxide: 1000µL of a stabilized 3% solution...Part 3012
Eosinophil Peroxidase: 100µL in 1 vial at 10 units/ml......Part 6016

The following kits are available:
Catalog	Size	Price (Eur)
FLEPO 100-2	100 Tests	€395

Figure 1. Eosinophil peroxidase was titrated in 1X reaction buffer and 50uL of each point was added in triplicate to 96 black wells. 50uL of reaction cocktail was added and the reaction was incubated in the dark at room temperature for 30 minutes.

Fluorescence was read at 530nm excitation and emission detected at 590nm.