Mpo activity assay kit

Manufactured by Abcam

Sourced in United States, United Kingdom

The MPO activity assay kit is a quantitative colorimetric assay designed to measure the activity of the enzyme myeloperoxidase (MPO) in biological samples. The kit provides reagents and a protocol to determine the MPO activity level in a straightforward and reliable manner.

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MPO activity (4 citations) Activity assay kits (2 citations) MPO kit (2 citations)

26 protocols using mpo activity assay kit

Quantifying Pancreatic and Pulmonary Inflammation

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Whole blood samples collected in anticoagulant treated tubes (450472, Greiner Bio-one) at necropsy were centrifuged at 2000g for 10 minutes at 4°C, and the resulting supernatant (serum) was collected for downstream analyses. Amylase activity in these mouse serum samples was measured commercially by IDEXX Bioanalytics using optimized protocol.
Trypsin enzymatic activity was measured from pancreas homogenates using trypsin activity assay kit (3043, Chondrex Inc.) following manufacturer’s instructions. Briefly, small pancreatic tissue pieces (25–30 mg) were washed in PBS, homogenized in 1× RIPA lysis buffer (ab156034, Abcam), and centrifuged at 13,000g for 10 minutes at 4°C. The total protein concentration from the collected supernatant was measured by BCA analysis (23225, Pierce Biotechnology), which was then used for the trypsin activity assay.
Neutrophil infiltration in the pancreas and lungs was quantified by measuring the MPO activity in the tissues utilizing the MPO Activity Assay Kit (K747, BioVision). For this assay, tissue samples (~20 mg) washed in PBS were homogenized in 500 μL of MPO Assay Buffer at 10,000g for 10 minutes at 4°C. Concentration of total protein from the collected supernatant was estimated by BCA analysis (23225, Pierce Biotechnology) and then utilized to measure MPO activity following manufacturer protocol.

Dey S., Udari L.M., RiveraHernandez P., Kwon J.J., Willis B., Easler J.J., Fogel E.L., Pandol S, & Kota J. (2021). Loss of miR-29a/b1 promotes inflammation and fibrosis in acute pancreatitis. JCI Insight, 6(19), e149539.

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Measuring Neutrophil Myeloperoxidase Activity

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Myeloperoxidase (MPO) is the functional and activating marker of neutrophils, and its level and activity change represent the function and active state of neutrophil polymorphonuclear leukocytes (PMN). The lung tissues were collected to measure MPO activity according to the manufacturer's instructions of the MPO Activity Assay Kit (K747-100, BioVision (Milpitas, CA, USA).

Wang J., Chen M., Xu W., Shou L., Jin X., Xu X, & Huang F. (2022). Inhibition of miR-29b-1-5p Attenuates Inflammatory Response and Pulmonary Fibrosis in LPS-Induced Acute Lung Injury by Regulating RTN4 Expression. Evidence-based Complementary and Alternative Medicine : eCAM, 2022, 7523591.

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Neutrophil Accumulation Quantification

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Myeloperoxidase (MPO) activity was evaluated as an index of neutrophil accumulation in the tissues by using the MPO Activity Assay Kit (BioVision) following the manufacturer’s protocol.

Dong D., Du Y., Fei X., Yang H., Li X., Yang X., Ma J., Huang S., Ma Z., Zheng J., Chan D.W., Shi L., Li Y., Irving A.T., Yuan X., Liu X., Ni P., Hu Y., Meng G., Peng Y., Sadler A, & Xu D. (2023). Inflammasome activity is controlled by ZBTB16-dependent SUMOylation of ASC. Nature Communications, 14, 8465.

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Liver Enzyme and Oxidative Stress Assays

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Alanine aminotransferase (ALT), aspartate aminotransferase (AST), myeloperoxidase (MPO), and catalase (CAT) activities and the malondialdehyde (MDA) concentration were measured using an ALT activity assay kit, AST activity assay kit, MPO activity assay kit, catalase assay kit, and lipid peroxidation (MDA) assay kit (Biovision, Milpitas, CA, USA). Ammonia concentration was measured using an ammonia assay kit (Abcam, Cambridge, UK).

Kwon K.W., Nam Y., Choi W.S., Kim T.W., Kim G.M, & Sohn U.D. (2019). Hepatoprotective effect of sodium hydrosulfide on hepatic encephalopathy in rats. The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology, 23(4), 263-270.

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Quantifying Lung Inflammation in Mice

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Mice were anesthetized with isoflurane by using the anesthesia system (R550, RWD Life Science) and euthanized with overdose CO₂. The analysis of immune cell infiltration and MPO (myeloperoxidase) activities was performed as described [31 (link)]. Briefly, CD45⁺ lymphocytes and neutrophils (CD45⁺CD11b⁺Ly-6G⁺Ly-6C^med) in BAL (bronchoalveolar lavage) were measured by flow cytometry. The MPO activity was determined by the MPO Activity Assay Kit (Abcam). HE (hematoxylin and eosin) staining of the lung tissue was performed as described [8 (link), 31 (link)].

Song J., Ma Q., Li Y., Wang X., Chen S., Liang B., Lin X., Chen J., Xu S., Shi S., Zhang J., Diao L., Zeng Y, & Xu J. (2024). CD317+ MSCs expanded with chemically defined media have enhanced immunological anti-inflammatory activities. Stem Cell Research & Therapy, 15, 2.

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Cardiac Injury Biomarkers and Cytokine Profiling

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After 24 hours of reperfusion, venous blood from mice was collected through the posterior orbital venous plexus to determine the serum troponin T level using an ELISA kit for troponin T (Mito Sciences). Next, the mice heart tissues were harvested to detect IFN‐γ (ab252352; Abcam), TNF‐α (ab208348; Abcam), IL‐4 (ab221833; Abcam) and IL‐13 (ab219634; Abcam) protein levels using an ELISA kit. The myeloperoxidase (MPO) level was detected using an MPO activity assay kit (ab105136; Abcam).

Yan M., Yang Y., Zhou Y., Yu C., Li R., Gong W, & Zheng J. (2021). Interleukin‐7 aggravates myocardial ischaemia/reperfusion injury by regulating macrophage infiltration and polarization. Journal of Cellular and Molecular Medicine, 25(21), 9939-9952.

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Measuring MPO Activity in EVs

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The MPO activity in EV samples and post-ultracentrifugation supernatants was measured with MPO activity assay kit (Abcam, ab105136) following the standard manual. The absorbance at 412 nm was measured using a microplate reader (SpectraMAX Gemini EM, Molecular Devices).

Iliev D.B., Strandskog G., Sobhkhez M., Bruun J.A, & Jørgensen J.B. (2021). Secretome Profiling of Atlantic Salmon Head Kidney Leukocytes Highlights the Role of Phagocytes in the Immune Response to Soluble β-Glucan. Frontiers in Immunology, 12, 736964.

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Murine Myeloperoxidase Activity Assay

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Murine MPO activity (muMPO) was detected by MPO activity assay kit (Abcam). Briefly, the samples were reacted with the reagents provided by the kit according to the manufacturer’s protocol. The samples were finally analyzed with a microplate reader at 412 nm.

An X.N., Wei Z.N., Xie Y.Y., Xu J., Shen Y., Ni L.Y., Shi H., Shen P.Y., Zhang W, & Chen Y.X. (2022). CD206+CD68+ mono-macrophages and serum soluble CD206 level are increased in antineutrophil cytoplasmic antibodies associated glomerulonephritis. BMC Immunology, 23, 55.

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Liver Tissue MPO Activity Assay

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Liver tissue MPO activity was measured by MPO Activity Assay Kit (Abcam, Cambridge, MA) according to the manufacturer’s instruction.

Xu J., Xue Z., Zhang C., Liu Y., Busuttil R.W., Zhang J., Kupiec-Weglinski J.W, & Ji H. (2019). Inhibition of Cyclin-dependent Kinase 2 Signaling Prevents Liver Ischemia and Reperfusion Injury. Transplantation, 103(4), 724-732.

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Serum Biomarker Quantification Techniques

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The serum levels of 8-OHdG, HEL, and PRL were measured using an on-chip ELISA,^{(29 (link))} and IL-6 levels in the serum were measured using a chemiluminescent enzyme immunoassay (CLEIA; LSI Medience Corporation, Tokyo, Japan). The plasma MPO levels were measured using an MPO activity assay kit (Abcam, Cambridge, UK).

Takeuchi K., Kazumura K., Yoshida A., Furuta T., Hayashi K., Nagai M., Hatano Y., Naito M, & Matsushita E. (2023). Comparison of the oxidative profiles before and after revascularization in peripheral arterial disease: a pilot study. Journal of Clinical Biochemistry and Nutrition, 72(3), 278-288.

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