Recombinant Mouse Myeloperoxidase Protein, CF

Catalog # Availability Size / Price Qty
3667-MP-250
Product Details
Citations (3)
FAQs
Supplemental Products
Reviews (1)

Recombinant Mouse Myeloperoxidase Protein, CF Summary

Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to oxidize guaiacol in the presence of hydrogen peroxide. Capeillere-Blandin, C. (1998) Biochem J. 336 :395. The specific activity is >8,000 pmol/min/μg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived mouse Myeloperoxidase/MPO protein
Met16-Thr718, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Met16
Predicted Molecular Mass
81 kDa
SDS-PAGE
85-95 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Assay Procedure

Materials
  • 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
  1. Prepare the substrate mixture by diluting guaiacol to 100 mM in Assay Buffer containing 0.0034% H2O2.
  2. Shake or stir for 15 minutes at room temperature. Protect from light.
  3. Dilute rmMPO to 3.34 µg/mL in Assay Buffer.
  4. 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).
  5. Read each cuvette at 470 nm in kinetic mode for 1 minute. Use only the first 10 seconds of data in the activity calculation.
  6. 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

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

References
  1. Klebanoff, S.J. (2005) J. Leukoc. Biol. 77:598.
  2. Hansson, M. et al. (2006) Arch. Biochem. Biophys. 445:214.
  3. Hashinaka, K. et al. (1988) Biochemistry 27:5906.
  4. van Dalen, C.J. et al. (2000) J. Biol. Chem. 275:11638.
  5. Hazen, S.L. and J.W. Heinecke (1997) J. Clin. Invest. 99:2075.
  6. Heinecke, J.W. et al. (1993) J. Clin. Invest. 91:2866.
  7. Podrez, E.A. et al. (1999) J. Clin. Invest. 103:1547.
  8. Bergt, C. et al. (2004) Proc. Natl. Acad. Sci. 101:13032.
  9. Hazen, S.L. et al. (1996) J. Biol. Chem. 271:23080.
  10. Eiserich, J.P. et al. (2002) Science 296:2391.
  11. Tiruppathi, C. et al. (2004) Proc. Natl. Acad. Sci. 101:7699.
  12. Aratani Y. et al. (2000) J. Infect. Dis. 182:1276.
  13. Kutter, D. (1998) J. Mol. Med. 76:669.
  14.  
Entrez Gene IDs
4353 (Human); 17523 (Mouse); 303413 (Rat)
Alternate Names
EC 1.11.1; EC 1.11.1.7; Lactoperoxidase; MPO; Myeloperoxidase

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.

3 Citations: Showing 1 - 3
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  1. 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;3(11):.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  2. 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;0(0):.
    Species: Bacteria - Staphylococcus aureus
    Sample Types: Cell Culture Supernates
    Applications: Bioassay
  3. 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;19(4):500-5.
    Species: N/A
    Sample Types: Protein
    Applications: Bioassay

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Recombinant Mouse Myeloperoxidase Protein, CF
By Anonymous on 05/10/2018
Application: Western