Recombinant Mouse Myeloperoxidase Protein, CF Summary
Product Specifications
Met16-Thr718, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
3667-MP
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS. |
| Reconstitution | Reconstitute at 500 μg/mL in sterile, deionized water. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Assay Procedure
- Assay Buffer: 20 mM MOPS, 0.1 M NaCl, 1 mM CaCl2, pH 7.0
- Recombinant Mouse Myeloperoxidase/MPO (rmMPO) (Catalog # 3667-MP)
- Hydrogen Peroxide Solution, 30% (w/w) (H2O2) (Sigma, Catalog # H1009)
- Guaiacol (Acros Organics, Catalog # AC120192500)
- Quartz Cuvette (Starna Cells, Catalog # 9B-Q-10) or equivalent
- Spectrophotometer with cuvette port (Model: Spectramax Plus by Molecular Devices) or equivalent
- Prepare the substrate mixture by diluting guaiacol to 100 mM in Assay Buffer containing 0.0034% H2O2.
- Shake or stir for 15 minutes at room temperature. Protect from light.
- Dilute rmMPO to 3.34 µg/mL in Assay Buffer.
- Load into a quartz cuvette 400 µL of 3.34 µg/mL rmMPO and start the reaction by adding 400 µL of the diluted guaiacol/H2O2 mixture. As a Substrate Blank combine 400 µL of Assay Buffer and 400 µL of the diluted guaiacol/H2O2 mixture (note: it is essential to monitor the reaction immediately after the introduction of the substrate mixture).
- Read each cuvette at 470 nm in kinetic mode for 1 minute. Use only the first 10 seconds of data in the activity calculation.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Absorbance change per minute ( delta A/min) x sample volume (L) x 1012 pmol/mol |
| ext. coeff (M-1cm-1) x amount of enzyme (µg) |
Notes:
Absorbance readings are adjusted for the Substrate Blank
Use an extinction coefficient of 5580 M-1cm-1
The output of many spectrophotometers is in milli absorbance units per minute in kinetic mode Per Reaction:
- rmMPO: 1.336 μg (20 nM)
- H2O2: 0.0017% (0.5 mM)
- Guaiacol: 50 mM
Reconstitution Calculator
Background: Myeloperoxidase/MPO
Myeloperoxidase (MPO) is a heme‑containing enzyme belonging to the XPO subfamily of peroxidases. It is an abundant neutrophil and monocyte glycoprotein that catalyzes the hydrogen peroxide‑dependent conversion of chloride, bromide, and iodide to multiple reactive species (1). Post‑translational processing of human MPO involves the insertion of a heme moiety and the proteolytic removal of both a propeptide and a 6 aa internal peptide (2). This results in a disulfide‑linked dimer composed of a 60 kDa heavy and 12 kDa light chain that associate into a 150 kDa enzymatically active tetramer. The tetramer contains two heme groups and one disulfide bond between the heavy chains (2). Mouse and human MPO share 87% aa sequence identity. MPO activity results in protein nitrosylation and the formation of 3‑chlorotyrosine and dityrosine crosslinks (4‑6). Modification of ApoB100, as well as the lipid and cholesterol components of LDL and HDL, promotes the development of atherosclerosis (5, 7‑9). MPO is also associated with a variety of other diseases (1), and inhibits vasodilation in inflammation by depleting the levels of NO (10). Serum albumin functions as a carrier protein during MPO movement to the basolateral side of epithelial cells (11). MPO is stored in neutrophil azurophilic granules. Upon cellular activation, it is deposited into pathogen‑containing phagosomes (2). While mice lacking MPO are impaired in clearing select microbial infections, MPO deficiency in humans does not necessarily result in heightened susceptibility to infections (12, 13).
- Klebanoff, S.J. (2005) J. Leukoc. Biol. 77:598.
- Hansson, M. et al. (2006) Arch. Biochem. Biophys. 445:214.
- Hashinaka, K. et al. (1988) Biochemistry 27:5906.
- van Dalen, C.J. et al. (2000) J. Biol. Chem. 275:11638.
- Hazen, S.L. and J.W. Heinecke (1997) J. Clin. Invest. 99:2075.
- Heinecke, J.W. et al. (1993) J. Clin. Invest. 91:2866.
- Podrez, E.A. et al. (1999) J. Clin. Invest. 103:1547.
- Bergt, C. et al. (2004) Proc. Natl. Acad. Sci. 101:13032.
- Hazen, S.L. et al. (1996) J. Biol. Chem. 271:23080.
- Eiserich, J.P. et al. (2002) Science 296:2391.
- Tiruppathi, C. et al. (2004) Proc. Natl. Acad. Sci. 101:7699.
- Aratani Y. et al. (2000) J. Infect. Dis. 182:1276.
- Kutter, D. (1998) J. Mol. Med. 76:669.
Citations for Recombinant Mouse Myeloperoxidase Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 5
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Eosinophils preserve bone homeostasis by inhibiting excessive osteoclast formation and activity via eosinophil peroxidase
Authors: Andreev, D;Kachler, K;Liu, M;Chen, Z;Krishnacoumar, B;Ringer, M;Frey, S;Krönke, G;Voehringer, D;Schett, G;Bozec, A;
Nature communications
Species: Murine polyomavirus strain A3
Sample Types: Whole Cells
Applications: Bioassay -
CD206<sup>+</sup>CD68<sup>+</sup> mono-macrophages and serum soluble CD206 level are increased in antineutrophil cytoplasmic antibodies associated glomerulonephritis
Authors: XN An, ZN Wei, YY Xie, J Xu, Y Shen, LY Ni, H Shi, PY Shen, W Zhang, YX Chen
BMC immunology, 2022-11-15;23(1):55.
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo -
Myeloperoxidase aggravates pulmonary arterial hypertension by activation of vascular Rho-kinase
Authors: A Klinke, E Berghausen, K Friedrichs, S Molz, D Lau, L Remane, M Berlin, C Kaltwasser, M Adam, D Mehrkens, M Mollenhaue, K Manchanda, T Ravekes, GA Heresi, M Aytekin, RA Dweik, JK Hennigs, L Kubala, E Michaëlsso, S Rosenkranz, TK Rudolph, SL Hazen, H Klose, RT Schermuly, V Rudolph, S Baldus
JCI Insight, 2018-06-07;3(11):.
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo -
Immune evasion by a staphylococcal inhibitor of myeloperoxidase
Authors: NWM de Jong, KX Ramyar, FE Guerra, R Nijland, C Fevre, JM Voyich, AJ McCarthy, BL Garcia, KPM van Kessel, JAG van Strijp, BV Geisbrecht, PA Haas
Proc. Natl. Acad. Sci. U.S.A., 2017-08-14;0(0):.
Species: Bacteria - Staphylococcus aureus
Sample Types: Cell Culture Supernates, Protein
Applications: Bioassay -
Enhanced detection of myeloperoxidase activity in deep tissues through luminescent excitation of near-infrared nanoparticles.
Authors: Zhang, Ning, Francis, Kevin P, Prakash, Arun, Ansaldi, Daniel
Nat Med, 2013-03-03;19(4):500-5.
Applications: Bioassay
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