Recombinant Human MMP-2 (NS0-expressed) Protein, CF

R&D Systems | Catalog # 902-MPN

R&D Systems
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Key Product Details

  • R&D Systems NS0-derived Recombinant Human MMP-2 (NS0-expressed) Protein (902-MPN)
  • 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

Applications

Enzyme Activity
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Product Specifications

Source

Mouse myeloma cell line, NS0-derived human MMP-2 protein
Ala30-Cys660

Purity

>90%, 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

Ala30

Predicted Molecular Mass

71 kDa (proform)

SDS-PAGE

60-70 kDa, 30 kDa, 21 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001).
The specific activity is >1,000 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

902-MPN
Formulation Lyophilized from a 0.2 μm filtered solution in Tris, CaCl2, NaCl and ZnCl2.
Reconstitution Reconstitute at 100 μg/mL in sterile 100 mM Tris, 10 mM CaCl2, 150 mM NaCl, and 0.05% Brij-35, pH 8.0.
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, -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-2

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‑2 (gelatinase A), a type IV collagenase, can degrade a broad range of substrates including type IV, V, VII and X collagens as well as elastin and fibronectin. It is believed to act synergistically with interstitial collagenase (MMP‑1) in the degradation of fibrillar collagens as it degrades their denatured gelatin forms. MMP‑2 has been shown to be associated with many connective tissue cells as well as neutrophils, macrophages and monocytes. Structurally, MMP‑2 may be divided into several distinct domains: a pro-domain which is cleaved upon activation; a catalytic domain containing the zinc binding site; a fibronectin-like domain thought to play a role in substrate targeting; and a carboxyl terminal (hemopexin-like) domain containing 2 N-linked glycosylation sites.

Long Name

Matrix Metalloproteinase 2

Alternate Names

Gelatinase A, MMP2

Entrez Gene IDs

4313 (Human); 17390 (Mouse)

Gene Symbol

MMP2

UniProt

Additional MMP-2 Products

Product Documents for Recombinant Human MMP-2 (NS0-expressed) 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.

Product Specific Notices for Recombinant Human MMP-2 (NS0-expressed) Protein, CF

For research use only

Citations for Recombinant Human MMP-2 (NS0-expressed) Protein, CF

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Protocols

View specific protocols for Recombinant Human MMP-2 (NS0-expressed) Protein, CF (902-MPN):

Materials
  • Activation Buffer: 100 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 8.0
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij 35, pH 7.5 (TCNB)
  • Recombinant Human MMP-2 (rhMMP-2) (Catalog # 902-MPN)
  • p-aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563), prepare a 100 mM stock in DMSO
  • Fluorogenic Peptide Substrate I: MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES001), 1 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhMMP-2 to 100 µg/mL in Activation Buffer.
  2. Activate rhMMP-2 by adding APMA to a final concentration of 1 mM.
  3. Incubate at 37 °C for 1 hour.
  4. Dilute activated rhMMP-2 to 0.2 ng/µL in assay buffer.
  5. Dilute substrate to 20 µM in assay buffer.
  6. Load in plate 50 µL of the 0.2 ng/µL rhMMP-2 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 substrate.
  7. Read at excitation and emission wavelengths of 320 nm and 405 nm, 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-2: 0.010 µg
  • Substrate: 10 µM

FAQs for Recombinant Human MMP-2 (NS0-expressed) Protein, CF

Showing  1 - 5 of 5 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 enzyme have a tag?

    A: No, this enzyme does not have a tag.  Please refer to the Source section on the product-specific page or product datasheet for sequence information.

  • 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 enzyme have a tag?

    A: No, this enzyme does not have a tag.  Please refer to the Source section on the product-specific page or product datasheet for sequence information.

  • 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 enzyme have a tag?

    A: No, this enzyme does not have a tag.  Please refer to the Source section on the product-specific page or product datasheet for sequence information.

  • 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 enzyme have a tag?

    A: No, this enzyme does not have a tag.  Please refer to the Source section on the product-specific page or product datasheet for sequence information.

  • 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 enzyme have a tag?

    A: No, this enzyme does not have a tag.  Please refer to the Source section on the product-specific page or product datasheet for sequence information.

  • 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 - 5 of 5 FAQs Showing All
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