Recombinant Human MMP-3 Protein, CF

R&D Systems | Catalog # 513-MP

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human MMP-3 Protein (513-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

P08254

Structure / Form

Pro form

Applications

Enzyme Activity
View all MMP-3 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human MMP-3 protein
Tyr18-Cys477 (Lys45Glu)

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Tyr18

Predicted Molecular Mass

52 kDa

SDS-PAGE

54-56 kDa doublet, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >150 pmol/min/µg, as measured under the described conditions.

Reviewed Applications

Read 3 reviews rated 5 using 513-MP in the following applications:

Formulation, Preparation, and Storage

513-MP
Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl, CaCl2, Glycerol and Brij-35.
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 opening.

Background: MMP-3

Matrix metalloproteinases are a family of zinc and calcium dependent endopeptidases with the combined ability to degrade all the components of the extracellular matrix. MMP-3 (stromelysin-1), can degrade a broad range of substrates including collagen alpha  chains, aggrecan, laminin, fibronectin, elastin, casein, alpha -1 antitrypsin, myelin basic protein, IL-1 beta, IGFBP-3, pro MMP-1, pro MMP-7, pro MMP-8, pro MMP-9 and pro MMP-13. MMP-3 does not cleave the triple helical region of interstitial collagens, a characteristic which distinguishes the stromelysins from the collagenases. The MMP-3 substrate repertoire extends beyond extracellular matrix proteins and implicates MMP-3 in roles other than direct tissue remodelling, for instance, enzyme cascades and cytokine regulation. MMP-3 is expressed by fibroblasts, chrondrocytes, osteoblasts, endothelial cells, smooth muscle cells and macrophages. Structurally, MMP-3 may be divided into several distinct domains; a pro-domain which is cleaved upon activation; a catalytic domain containing the zinc binding site; a short hinge region and a carboxyl terminal (hemopexin-like) domain.

Long Name

Matrix Metalloproteinase 3

Alternate Names

MMP3, Stromelysin 1

Entrez Gene IDs

4314 (Human); 17392 (Mouse)

Gene Symbol

MMP3

UniProt

P08254

Additional MMP-3 Products

Product Documents for Recombinant Human MMP-3 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.

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Product Specific Notices for Recombinant Human MMP-3 Protein, CF

For research use only

Citations for Recombinant Human MMP-3 Protein, CF

Customer Reviews for Recombinant Human MMP-3 Protein, CF (3)

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  • Recombinant Human MMP-3 Protein, CF
    Name: Andres Ocampo Carrillo
    Application: In vivo study
    Verified Customer | Posted 03/04/2023
    rhMMP-3 protein was dissolved, stored as indicated, aliquoted, and used several times to induce pro-inflammatory responses in an in vivo model of intestinal inflammation. Application of this protein induced a strong accumulation of cells in the colon of the animals used (CD-1/ICR mice, treatment: 7 days, 10 ng/mL).
    Recombinant Human MMP-3 Protein, CF 513-MP
  • Recombinant Human MMP-3 Protein, CF
    Name: Anonymous
    Application: Binding assay/Protein-protein interaction
    Verified Customer | Posted 03/22/2018
  • Recombinant Human MMP-3 Protein, CF
    Name: Anonymous
    Application: Immunoassay Standard
    Verified Customer | Posted 11/10/2017
    The recombinant MMP-3 protein was used as to build an ELISA standard for an assay designed to measure MMP-3 in human serum and plasma samples.
    Recombinant Human MMP-3 Protein, CF 513-MP

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Protocols

View specific protocols for Recombinant Human MMP-3 Protein, CF (513-MP):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Human MMP-3 (rhMMP-3) (Catalog # 513-MP)
  • Chymotrypsin (Sigma, Catalog # C-3142), 1 mg/mL stock in 1 mM HCl
  • Phenylmethyl Sulfonyl Fluoride (PMSF) (Sigma, Catalog # P-7626), 0.2 M stock in 2-Propanol
  • Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002), 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Activate rhMMP-3 at 20 µg/mL in Assay Buffer containing 5 µg/mL Chymotrypsin.
  2. Incubate reaction at 37 °C for 30 minutes.
  3. Stop activation with 2 mM PMSF. Pre-warm the PMSF to 37 °C prior to adding to sample.
  4. Dilute activated rhMMP-3 to 2.5 ng/µL in Assay Buffer.
  5. Dilute Substrate to 20 µM in Assay Buffer.
  6. In a plate load 50 µL of 2.5 ng/µL rhMMP-3, and start the reaction by adding 50 µL of 20 µM Substrate to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
  7. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  8. 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:

  • rhMMP-3: 0.125 µg
  • Substrate: 10 µM

FAQs for Recombinant Human MMP-3 Protein, CF

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    • 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.

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

    • 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.
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