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Recombinant Human MMP-9 Protein, CF

R&D Systems | Catalog # 911-MP

Analyzed by SEC-MALS
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Key Product Details

  • R&D Systems CHO-derived Recombinant Human MMP-9 Protein (911-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

CHO

Accession Number

P14780

Structure / Form

Pro form

Applications

Enzyme Activity
View all MMP-9 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human MMP-9 protein
Ala20-Asp707 (Gln279Arg)

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.1 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ala20

Predicted Molecular Mass

77 kDa

SDS-PAGE

93 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 >1,300 pmol/min/µg, as measured under the described conditions.

Reviewed Applications

Read 8 reviews rated 4.6 using 911-MP in the following applications:

Scientific Data Images for Recombinant Human MMP-9 Protein, CF

Recombinant Human MMP‑9 Protein SEC-MALS.

Recombinant Human MMP‑9 Protein (Catalog # 911-MP) has a molecular weight (MW) of 88-97 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
Recombinant Human MMP-9 Protein Enzyme Activity

Recombinant Human MMP-9 Protein Enzyme Activity

Recombinant Human MMP-9 (Catalog # 911-MP) is measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001).

Formulation, Preparation, and Storage

911-MP
Formulation Supplied as a 0.2 μm filtered solution in Tris, CaCl2, NaCl 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-9

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-9 (gelatinase B) can degrade a broad range of substrates including gelatin, collagen types IV and V, elastin and proteoglycan core protein. 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-9 is produced by keratinocytes, monocytes, macrophages and PMN leukocytes. MMP-9 is present in most cases of inflammatory responses. Structurally, MMP-9 maybe be divided into five distinct domains: a pro-domain which is cleaved upon activation, a gelatin-binding domain consisting of three contiguous fibronectin type II units, a catalytic domain containing the zinc binding site, a proline-rich linker region, and a carboxyl terminal hemopexin-like domain. In addition to the human enzyme, the recombinant mouse MMP-9 is also available (Catalog # http://www.rndsystems.com/product_results.aspx?k=909-MM">909-MM).

Long Name

Matrix Metalloproteinase 9

Alternate Names

CLG4B, Gelatinase B, GELB, MANDP2, MMP9

Entrez Gene IDs

4318 (Human); 17395 (Mouse); 81687 (Rat); 102117693 (Cynomolgus Monkey)

Gene Symbol

MMP9

UniProt

P14780

Additional MMP-9 Products

Product Documents for Recombinant Human MMP-9 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 Human MMP-9 Protein, CF

For research use only

Citations for Recombinant Human MMP-9 Protein, CF

Customer Reviews for Recombinant Human MMP-9 Protein, CF (8)

4.6 out of 5
8 Customer Ratings
5 Stars
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4 Stars
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3 Stars
13%
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1 Stars
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  • Recombinant Human MMP-9 Protein, CF
    Name: Jan Van Deun
    Application: Enzymatic activity in vitro
    Verified Customer | Posted 08/04/2023
    Incubating 1h at 37°C after addition of APMA was sufficient for activation.
    Recombinant Human MMP-9 Protein, CF 911-MP
  • Recombinant Human MMP-9 Protein, CF
    Name: Andres Ocampo Carrillo
    Application: In vitro bioactivity in cell culture
    Verified Customer | Posted 03/04/2023
    rhMMP-9 was used to treat human chondrocytes to induce an enzymatic cleavage of chondrocytes extracellular proteins and produce several prolactin fragments. A comparison with several MMP’s showed a successful generation of fragments at concentrations ranging from 10 to 10000 ng, indicating the validity of the enzymatic activity of MMP-9
    Recombinant Human MMP-9 Protein, CF 911-MP
  • Recombinant Human MMP-9 Protein, CF
    Name: Oleg Yerov
    Application: Enzymatic activity in vitro
    Verified Customer | Posted 11/06/2020
  • Recombinant Human MMP-9 Protein, CF
    Name: Anonymous
    Application: Enzymatic activity in vitro
    Verified Customer | Posted 10/09/2019
  • Name: Anonymous
    Application: Enzymatic activity in vitro
    Verified Customer | Posted 08/14/2018
    Recombinant Human MMP-9 Protein, CF 911-MP
  • Recombinant Human MMP-9 Protein, CF
    Name: Radhika Mehta
    Application: Binding assay/Protein-protein interaction
    Verified Customer | Posted 03/22/2018
  • Name: Balaji Mahender
    Application: Immunoassay Standard
    Verified Customer | Posted 12/20/2017
  • Recombinant Human MMP-9 Protein, CF
    Name: Anonymous
    Application: Immunoassay Standard
    Verified Customer | Posted 11/10/2017
    The recombinant human MMP-9 protein was used to create an ELISA standard for an immunoassay to measure MMP-9 in serum samples.
    Recombinant Human MMP-9 Protein, CF 911-MP

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Protocols

View specific protocols for Recombinant Human MMP-9 Protein, CF (911-MP):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35 (w/v), pH 7.5 (TCNB)
  • Recombinant Human MMP-9 (rhMMP-9) (Catalog # 911-MP)
  • p-aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563), prepare a 100 mM stock in DMSO
  • Substrate: MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES001)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhMMP-9 to 100 µg/mL in Assay Buffer.
  2. Activate rhMMP-9 by adding APMA to a final concentration of 1 mM.
  3. Incubate at 37 °C for 24 hours.
  4. Dilute activated rhMMP-9 to 0.4 ng/µL in Assay Buffer.
  5. Dilute Substrate to 20 µM in Assay Buffer.
  6. Load 50 µL of the 0.4 ng/µL rhMMP-9 into a plate and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL of 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-9: 0.020 µg
  • Substrate: 10 µM

FAQs for Recombinant Human MMP-9 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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