Recombinant Mouse Myeloperoxidase Protein, CF

R&D Systems | Catalog # 3667-MP

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Myeloperoxidase Protein (3667-MP)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

AAR99349

Applications

Enzyme Activity
View all Myeloperoxidase/MPO Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Myeloperoxidase/MPO protein
Met16-Thr718, with a C-terminal 10-His tag

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.

N-terminal Sequence Analysis

Met16

Predicted Molecular Mass

81 kDa

SDS-PAGE

60-70 kDa and 85-95 kDa, reducing conditions

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.

Reviewed Applications

Read 2 reviews rated 4.5 using 3667-MP in the following applications:

Formulation, Preparation, and Storage

3667-MP
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution

Reconstitute at&nbsp;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.

Calculators

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.  

Alternate Names

MPO

Entrez Gene IDs

4353 (Human); 17523 (Mouse); 303413 (Rat)

Gene Symbol

MPO

UniProt

AAR99349

Additional Myeloperoxidase/MPO Products

Product Documents for Recombinant Mouse Myeloperoxidase Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse Myeloperoxidase Protein, CF

For research use only

Citations for Recombinant Mouse Myeloperoxidase Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Myeloperoxidase Protein, CF (3667-MP):

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

FAQs

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