Fluoro MPO

Product Description

Key Benefits

• Can monitor multiple time points to follow kinetics.
• One-step, no wash assay.
• Adaptable for High Throughput format.
• Highly Sensitive.
• Applications – Fluorescent Plate Reader or Absorbance Plate Reader.

Assay Principle

• Detection of MPO 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 oxidative tissue damage.
• MPO activity in acute and chronic manifestations of cardiovascular disease.

Figure 1. MPO standard curve was serially diluted in 1X Reaction Buffer. Reaction cocktail (RC) was prepared as described (without EPO inhibitor). Next 50uL of MPO standard and 50uL of RC was added to individual well of a 96 well black plates. The plate was incubated at room and temperature in the dark. Data collected Ex:530nm Em:590nm

Applications

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 powerfull bactericidal produced by neutrophils (4 review). Excessive production of these radicals can cause oxidative stress leading to oxidative tissue injury.

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

Excitation:530-571nm; Emission: 590-600nm

Citations

Use of a hanging-weight system for liver ischemic preconditioning in mice - Hart, Much, Kohler et.al - Am J Physiol Gastrointest Liver Physiol 294: G1431-G1440, 2008

Extracellular Adenosine Production by Ecto-5′-Nucleotidase Protects During Murine Hepatic Ischemic Preconditioning - M. Hart, C. Much, I. Gorzolla, J. Schittenhelm, D. Kloor, G. Stahl, H. Eltzschig - Gastroenterology Volume 135, Issue 5, Pages 1739-1750.e3

Inflammatory Biomarkers of Sulfur Mustard Analog 2-Chloroethyl Ethyl Sulfide–Induced Skin Injury in SKH-1 Hairless Mice - Neera Tewari-Singh, Sumeet Rana, Mallikarjuna Gu, Arttatrana Pal, David J. Orlicky, Carl W. White and Rajesh Agarwal - Toxicological Sciences 2009 108(1):194-206

References

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• Andrews, P.C., Parnes, C. & Krinsky, N.I. (1984) Comparison of myeloperoxidase and hemi-myeloperoxidase with respect to catalysis, regulation, and bacterial activity. Arch. Biochem. Biophys. 228, 439–442.
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Kit contents and Long Term storage