Recombinant Mouse MMP-12 Protein, CF

R&D Systems | Catalog # 3467-MPB

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse MMP-12 Protein (3467-MPB)
  • 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

EDL24933

Structure / Form

Pro form

Applications

Enzyme Activity
View all MMP-12 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse MMP-12 protein
Ala18-Cys462

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ala18

Predicted Molecular Mass

52 kDa

SDS-PAGE

50-62 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic peptide substrate Mca-KPLGL-Dpa-AR-NH2 (Catalog # ES010).
The specific activity is >100 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

3467-MPB
Formulation Lyophilized from a 0.2 μm filtered solution in MES, NaCl, CaCl2, CHAPS, ZnSO4 and PEG.
Reconstitution Reconstitute at 0.25 mg/mL in 10 mM MES, 0.1 M NaCl, 100 µM CaCl2, 0.1% CHAPS, 1 µM ZnSO4 and 0.1% PEG, pH 6.0
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: MMP-12

Matrix metalloproteinases (MMPs) are a family of zinc and calcium dependent endopeptidases that collectively degrade the components of the extracellular matrix. MMP-12 (macrophage elastase) consists of the following domains: a pro domain, a catalytic domain containing the zinc-binding site, and a C-terminal hemopexin-like domain. The 52 kDa rmMMP-12 pro form is activated via processing into 45 kDa and 22 kDa active forms (1). MMP-12 is capable of cleaving several substrates in addition to elastin. Macrophage secretion of MMP-12 at sites of inflammation can be induced by cytokines. Given the secretion of MMP12 during an inflammatory response, MMP-12 is involved in many pathologies including vascular disease (2, 3) and cancer (4-6). In particular, MMP12-mediated pathological degradation of the extracellular matrix is a well-established key event in inflammatory-related pulmonary disease (7). For example, overexpression of MMP12 in alveolar macrophages is associated with smoking and emphysema (8) while different MMP-12 variants result in protection against chronic obstructive pulmonary disease (COPD) development (9) or cause increased risk and disease severity (10). Recently, MMP-12 has been shown to translocate into the nucleus of viral-infected cells to directly regulate transcription during an immune response (11).

References

  1. Shapiro, S.D. et al. (1993) J. Biol. Chem. 268:23824.
  2. Liu, S.L. et al. (2015) Sci. Rep. 5:17189.
  3. Iyer, R.P. et al. (2015) Int. J. Cardiol. 185:198.
  4. Cao, W. et al. (2017) Oncol. Rep. 3:1401.
  5. Klupp, F. et al. (2016) BMC Cancer 16:494.
  6. Chung, I.C. et al. (2014) BMC Cancer 14:348.
  7. Vanderbroucke, R.E. et al. (2011) Eur. Respir. J. 387:1200.
  8. Babusyte, A. et al. (2007) Respir. Res. 8:81.
  9. Hunninghake, G.M. et al. (2009) N. Engl. J. Med. 361:2599.
  10. Mukhopadhyay, S. et al. (2010) J. Allergy Clin. Immunol. 126:70.
  11. Marchant, D.J. et al. (2013) Nature Medicine 20:493.

Long Name

Matrix Metalloproteinase 12

Alternate Names

MMP12

Entrez Gene IDs

4321 (Human); 17381 (Mouse)

Gene Symbol

MMP12

UniProt

EDL24933

Additional MMP-12 Products

Product Documents for Recombinant Mouse MMP-12 Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Mouse MMP-12 Protein, CF

For research use only

Related Research Areas

Metalloproteases and Regulators

Citations for Recombinant Mouse MMP-12 Protein, CF

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Protocols

View specific protocols for Recombinant Mouse MMP-12 Protein, CF (3467-MPB):

Materials
  • Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5
    (TCNB) + 5 µM ZnCl2
  • Assay  Buffer: 50 mM Tris, 10 mM CaCl2,150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant  Mouse MMP-12 (rmMMP-12) (Catalog # 3467-MPB)
  • p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A9563), 100 mM stock in DMSO
  • Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES010), 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmMMP-12 to 100 µg/mL with Activation Buffer containing 1 mM APMA.
  2. To activate, incubate rmMMP-12 at 37 °C for 24 hours.
  3. Dilute activated rmMMP-12 to 2 µg/mL in Assay Buffer.
  4. Dilute Substrate to 20 µM in Assay Buffer.
  5. Load 50 µL of 2 µg/mL rmMMP-12 to wells, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate blank containing 50 µL Assay Buffer and 50 µL of 20 µM.
  6. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. Calculate specific activity:
     

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank.                                                                                      

**Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:

  • rmMMP-12: 0.1 µg
  • Substrate: 10 µM

FAQs for Recombinant Mouse MMP-12 Protein, CF

Showing  1 - 4 of 4 FAQs Showing All

  • Q: Can the enzyme be stored after activation, or do I need to use it immediately after activation?

    A: We recommend only activating the amount of enzyme needed for your assay, and recommend activating the enzyme immediately prior to use. Any unactivated enzyme should be stored in aliquots at either the stock concentration at which the enzyme was supplied, or the reconstitution concentration, according to the product datasheet.

  • Q: Does this MMP enzyme need to be activated to work?

    A: Yes, this enzyme requires activation prior to use.

  • Q: If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?

    A: We have only optimized activation conditions for one particular concentration of this MMP enzyme as part of our regular QC testing for enzymatic activity. Activating the enzyme at any different concentration would have to be optimized by the end user.

  • Q: What is the activity of this enzyme in units/µg?

    A: We supply this enzyme as a mass and calculate its activity relative to mass (pmol/min/µg). We have not calibrated this enzyme to an international standard unit, so we are unable to provide a conversion to units/µg.

  • Q: Can the enzyme be stored after activation, or do I need to use it immediately after activation?

    A: We recommend only activating the amount of enzyme needed for your assay, and recommend activating the enzyme immediately prior to use. Any unactivated enzyme should be stored in aliquots at either the stock concentration at which the enzyme was supplied, or the reconstitution concentration, according to the product datasheet.

  • Q: Does this MMP enzyme need to be activated to work?

    A: Yes, this enzyme requires activation prior to use.

  • Q: If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?

    A: We have only optimized activation conditions for one particular concentration of this MMP enzyme as part of our regular QC testing for enzymatic activity. Activating the enzyme at any different concentration would have to be optimized by the end user.

  • Q: What is the activity of this enzyme in units/µg?

    A: We supply this enzyme as a mass and calculate its activity relative to mass (pmol/min/µg). We have not calibrated this enzyme to an international standard unit, so we are unable to provide a conversion to units/µg.

  • Q: Can the enzyme be stored after activation, or do I need to use it immediately after activation?

    A: We recommend only activating the amount of enzyme needed for your assay, and recommend activating the enzyme immediately prior to use. Any unactivated enzyme should be stored in aliquots at either the stock concentration at which the enzyme was supplied, or the reconstitution concentration, according to the product datasheet.

  • Q: Does this MMP enzyme need to be activated to work?

    A: Yes, this enzyme requires activation prior to use.

  • Q: If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?

    A: We have only optimized activation conditions for one particular concentration of this MMP enzyme as part of our regular QC testing for enzymatic activity. Activating the enzyme at any different concentration would have to be optimized by the end user.

  • Q: What is the activity of this enzyme in units/µg?

    A: We supply this enzyme as a mass and calculate its activity relative to mass (pmol/min/µg). We have not calibrated this enzyme to an international standard unit, so we are unable to provide a conversion to units/µg.

  • Q: Can the enzyme be stored after activation, or do I need to use it immediately after activation?

    A: We recommend only activating the amount of enzyme needed for your assay, and recommend activating the enzyme immediately prior to use. Any unactivated enzyme should be stored in aliquots at either the stock concentration at which the enzyme was supplied, or the reconstitution concentration, according to the product datasheet.

  • Q: Does this MMP enzyme need to be activated to work?

    A: Yes, this enzyme requires activation prior to use.

  • Q: If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?

    A: We have only optimized activation conditions for one particular concentration of this MMP enzyme as part of our regular QC testing for enzymatic activity. Activating the enzyme at any different concentration would have to be optimized by the end user.

  • Q: What is the activity of this enzyme in units/µg?

    A: We supply this enzyme as a mass and calculate its activity relative to mass (pmol/min/µg). We have not calibrated this enzyme to an international standard unit, so we are unable to provide a conversion to units/µg.

Showing  1 - 4 of 4 FAQs Showing All
View all FAQs for Proteins and Enzymes