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| Sample Type(s) | Cell lysates, tissues, etc |
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Overview
For quantitative determination of myeloperoxidase enzyme activity and evaluation of drug modulators. The assay uses FL530/585nm for signal readout. Compatible sample input includes Cell lysates, tissues, etc. Typical stated assay timing is Approximately 30 min.
Key elements and design rationale
- Readout format: FL530/585nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Cell lysates, tissues, etc, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 0.0025 U/L for interpreting low-signal samples.
- Feature emphasis: Fast and sensitive. Linear detection range (20 µL sample): 0.0025 to 2 U/L for 10 min reaction at 25°C.
Additional feature notes highlight Convenient and high-throughput. Homogeneous “mix-incubate-measure” type assay. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of enzyfluotm myeloperoxidase within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
MYELOPEROXIDASE (MPO; EC 1.11.2.2) is a peroxidase enzyme and can be found in neutrophils, monocytes, and some soft tissue macrophages. MPO has the ability to use chloride as a cosubstrate with hydrogen peroxide to generate hypochlorous acid, a powerful antimicrobial agent produced by neutrophils. However, excessive production of hypochlorous acid can lead to oxidative stress and tissue damage. Inflammation may also result when MPO oxidizes various substances such as phenols and anilines. Studies show that increased MPO levels may increase the risk of myocardial infarction and cardiovascular disease.BioAssay Systems EnzyFluoTMmyeloperoxidase (MPO) assay kit is based on the MPO enzyme reaction with hydrogen peroxide (H2O2) which oxidizes the dye reagent to a highly fluorescent product. The fluorescence intensity of this product, measured at &labda; ex/em = 530/585 nm, is proportional to the total peroxidation activity in the sample. The provided MPO inhibitor is used to suppress peroxidase activity due to MPO in order to differentiate other peroxidase activities that may be present in the samples.Related Categories:Oxidative-Stress.htm” target=_blank>Oxidative Stress, Enzyme-Activity-Assays.htm” target=_blank>Enzyme Activity Assays
Detection method
Fluorescent (FL 530/585 nm).
Detection limit and analytical sensitivity
Reported detection limit: 0.0025 U/L.
Procedures and timing
Stated procedure or timing information: Approximately 30 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify enzyfluotm myeloperoxidase in cell lysates, tissues by FL530/585 nm readout.
- Compare treatment or phenotype groups using matched cell lysates, tissues handling.
- Monitor time-course or pre/post changes in cell lysates, tissues across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
What samples have you tested?
The assay kit has been tested in purified myeloperoxidase, tissue and cell samples. Please follow the directions on the protocol for sample pretreatment.
What is the benefit of using a myeloperoxidase inhibitor in the assay?
The provided MPO inhibitor is used to suppress peroxidase activity due to MPO in order to differentiate other peroxidase activities that may be present in the samples. The difference in fluorescence intensities between sample and this sample blank is used to compute enzyme activity specifically due to myeloperoxidase.
I don’t have the correct wavelength filter, what other wavelength(s) would work?
The assay can be measured at excitation 500-540 nm and emission at 560-590 nm. Reading outside this range will result in a significant decrease in sensitivity.
Will the assay kit work in a 384 well plate?
Yes, the assay can be used in any standard plate. Simply adjust the sample and reagent volumes accordingly. For 384 well use 20-90 µL final reaction volume.
Can I store unused reagents for future use?
Yes, unused reagents can be stored according to the assay protocol. Repeated freeze/thaw cycles of reagents should be avoided. Working Reagents should be made fresh for each assay and used within 1 hour.
Do I need to use a standard or standard curve with each assay run?
Yes, it is highly recommended run the resorufin standard in each assay.
For laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Ca2+ influx channel inhibitor saraf protects mice from acute pancreatitis
Son, A., et al (2019). Ca2+ influx channel inhibitor saraf protects mice from acute pancreatitis. Gastroenterology, 157(6), 1660-1672.e2. Assay: Myeloperoxidase in mouse tissue.
Cysteinyl leukotrienes predominantly mediate cisplatin-induced acute renal damage in male rats
Hashim, A. A., Helmy, M. M., & Mouneir, S. M. (2018). Cysteinyl leukotrienes predominantly mediate cisplatin-induced acute renal damage in male rats. Journal of Physiology and Pharmacology, 69(5), 779-787. Assay: Myeloperoxidase in rat kidney.
Zileuton alleviates acute cisplatin nephrotoxicity: Inhibition of lipoxygenase pathway favorably modulates the renal oxidative/inflammatory/caspase-3 axis
Helmy, M. M., Hashim, A. A., & Mouneir, S. M. (2018). Zileuton alleviates acute cisplatin nephrotoxicity: Inhibition of lipoxygenase pathway favorably modulates the renal oxidative/inflammatory/caspase-3 axis. Prostaglandins & other lipid mediators, 135, 1-10. Assay: Myeloperoxidase in rat tissue.
