Myeloperoxidase Activity Assay Kit

NWLSS | Northwest Life Science Specialties, LLC.
Myeloperoxidase (MPO) Activity Assay
NWK-MPO03 $530.00 each

For questions & ordering, call:
360-449-3091
(Toll-Free US/Canada)



A simple and easy colorimetric assay for the study of myeloperoxidase (MPO) activity. This assay is not species specific and is compatible in all model systems where myeloperoxidase enzyme is thought to be present.

Introduction

Myeloperoxiase (MPO), the most abundant protein in neutrophils (also found in monocytes), is the focus of inflammatory pathologies. Most recent work has indicated that it is an excellent biomarker for human cardiovascular risk.

Its ability to catalyze reaction between chloride and hydrogen peroxide (H2O2) to form hypochlorous acid is unique among mammalian enzymes and is considered to be the dominant function of MPO in vivo. Hypochlorous acid is a powerful antimicrobial agent, and extremely reactive with biological molecules causing much of the damage mediated by neutrophils in inflammatory diseases. MPO also exhibits peroxidase activity that catalyzes oxidation of a number of substrates by (H2O2). This activity has been widely used to assess the amount of MPO.

Unfortunately, its specificity is very poor for unpurified biological samples due to the presence of other peroxidases. Peroxidases however, generally do not produce hypochlorous acid; the only exception is eosinophil peroxidase that produces hypochlorous acid at pH levels below 5. The chlorination activity of MPO has a pH optinum of near neutral pH. Therefore, assay conditions can be set so to provide for MPO enzyme specificity.

Test Principle

This method has been described by Weiss and coworkers (1982). Briefly, hypochlorous acid (HOCl) is formed from MPO catalyzed reaction between chloride and hydrogen peroxide. HOCl is rapidly trapped by β-amino acid taurine to form a stable oxidant taurine chloramine. Taurine prevents accumulation of hypochlorous acid that could deactivate MPO and does not react with MPO enzyme intermediate to interfere MPO catalysis.

After incubation for specific time, the MPO catalyzed reaction is stopped by adding catalase to eliminate hydrogen peroxide. Taurine chloramine thus formed is then allowed to react with 5-thio-2-nitrobenzoic acid (TNB). TNB has a chromophore that has maximal absorbance at 412 nm while its reaction product with taurin chloramine, 5-5-dithiobis(2-nitrobenzoic acid) or DTNB is colorless.

By following the decrease of absorbance at 412 nm, MPO activity is measured. One unit is the amount of MPO that can produce 1.0 nmole of taurine chloramine (hypochlorous acid) at pH 6.5 and 25C during 30 minutes in the presence of 100 mM chloride and 100 mM of hydrogen peroxide. The assay principal is summarized in the reaction scheme below.

MPO03 Reaction

Since DTNB and taurine does not absorb at 412 nm, the absorbance of TNB at 412 nm is measured to calculate the MPO unit directly. In the presence of MPO, the decrease of absorbance at 412 nm is proportionally (linearly) enhanced.

Pulmonary exposure to cellulose nanocrystals caused deleterious effects to reproductive system in male mice
Mariana T. Farcas, Elena R. Kisin, Autumn L. Menas, Dmitriy W. Gutkin, Alexander Star, Richard S. Reiner, Naveena Yanamala, Kai Savolainen & Anna A. Shvedova
Journal of Toxicology and Environmental Health, Part A,  Volume 79, 2016 - Issue 21, Pages 984-997

View Abstract

Intravenous Pyruvate to Prevent Renal Injury Following Cardiac Arrest and Resuscitation
Roger A. Hollrah M.S
University of North Texas Health Science Center, UNTHSC Scholarly Repository ,  Dissertation, August 1, 2014

View Abstract

Alleviation of Inhibitory Effects of Staphylococcus aureus on MPO Expression and Myeloperoxidase Activity in Human Neutrophils in vitro by Piper betle Linn. Ethanolic Extract
Yalda Modirrahmati, Mohd Fahmi Mastuki and Roslinah Mohamad Hussain
Journal of Applied Sciences ,  2015; 15 (6): 874-883

View Abstract

Is myeloperoxidase level in ascites a predictive factor for spontaneous bacterial peritonitis?
Yüksel Seçkin, Yasir Furkan Çağın, Fatma Sener
J Turgut Ozal Med Cent.,  April 9, 2016, Available Online Ahead of Print

View Abstract

The Effects of Epidural Anesthesia on Growth of Escherichia coli at Pseudosurgical Site: The Roles of the Lipocalin-2 Pathway
Igarashi, Toru MD; Suzuki, Takeshi MD; Mori, Katsuya PhD; Inoue, Kei MD; Seki, Hiroyuki MD; Yamada, Takashige MD; Kosugi, Shizuko MD; Minamishima, Shizuka MD; Katori, Nobuyuki MD; Sano, Fumiya MS; Abe, Takayuki PhD; Morisaki, Hiroshi MD
Anesthesia & Analgesia July 2015; Vol. 121 - Issue 1: p 81–89

Link to Abstract

Browse all Citations

For Research Use Only

Catalog Number: NWK-MPO03
Format: Enzymatic
Sample Requirements: Plasma, tissue homogenates, and other biological fluids where MPO activity may be present
Specificity: Myeloperoxidase activity in all species
Sensitivity: LLD = 10U/mL in sample assayed
Storage and Stability: Six months from date of manufacture when stored at 2-8C
Kit Contents: Assay Buffer
H2O2 Reagent
Catalase Reagent
TNB Reagent
Concentrated Taurine Solution
Random Citation | NWK-MPO03

Exposure to polychlorinated biphenyls enhances lipid peroxidation in human normal peritoneal and adhesion fibroblasts: A potential role for myeloperoxidase 
Ghassan M. Saed, Zhong L. Jianga, Nicole M. Fletchera, Ali Al Araba, Michael P. Diamonda and Husam M. Abu-Soud 
Free Radical Biology and Medicine, Volume 48, Issue 6, 15 March 2010, Pages 845-850 

Link to Abstract

The cart is empty

Not sure if this is the right biomarker for your project?

Let us help! Give our specialists a call (or contact us online) and let us help you figure out the best biomarker for your needs. We love discussing research!

360-449-3091
(Mon-Fri, 8am-5pm PST)

For questions, orders, and technical service:

 
360-449-3091
(Direct)
 
360-449-3091
(Toll-Free US/Canada)
 
360-397-0181
(Fax)
Online Support