Recombinant Rat MMP-9 Protein, CF

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Summary
Reactivity RtSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Rat MMP-9 Protein, CF Summary

Details of Functionality
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001). The specific activity is >710 pmol/min/μg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived rat MMP-9 protein
Leu35-Pro708, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Leu35
Structure / Form
Pro form
Protein/Peptide Type
Recombinant Enzymes
Gene
Mmp9
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
75 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
94 kDa, reducing conditions
Reviewed Applications
Read 1 Review rated 5
using
5427-MM in the following application:

Publications
Read Publications using
5427-MM in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in MES, NaCl, CaCl2 and Glycerol.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Rat MMP-9 (rrMMP-9) (Catalog # 5427-MM)
  • p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A-9563), 100 mM stock in DMSO
  • Substrate: MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES001), 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. Dilute rrMMP-9 to 100 µg/mL in Assay Buffer.
  2. Activate rrMMP-9 by adding APMA to a final concentration of 1 mM.
  3. Incubate activation reaction at 37 °C for 8 hours.
  4. Dilute activated rrMMP-9 to 0.2 ng/µL in Assay Buffer.
  5. Dilute Substrate to 20 µM in Assay Buffer.
  6. In a plate load 50 µL of 0.2 ng/µL rrMMP-9, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank of 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:
  • rrMMP-9: 0.01 µg
  • Substrate: 10 µM
  • Notes

    This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

    Alternate Names for Recombinant Rat MMP-9 Protein, CF

    • 92 kDa gelatinase
    • 92 kDa type IV collagenase
    • CLG4B
    • EC 3.4.24
    • EC 3.4.24.35
    • Gelatinase B
    • GELB
    • macrophage gelatinase
    • MANDP2
    • matrix metallopeptidase 9
    • matrix metalloproteinase 9
    • matrix metalloproteinase-9
    • MMP9
    • MMP-9
    • type V collagenase

    Background

    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 may be divided into five distinct domains: a pro-domain which is cleaved upon activation, a catalytic domain containing the zinc binding site, a gelatin-binding domain consisting of three contiguous fibronectin type II units, a proline-rich linker region, and four carboxyl terminal hemopexin-like domains.

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    Publications for MMP-9 (5427-MM)(4)

    We have publications tested in 2 confirmed species: Mouse, Rat.

    We have publications tested in 3 applications: Spectrum Measurement, Western Blot, Zymography.


    Filter By Application
    Spectrum Measurement
    (1)
    Western Blot
    (1)
    Zymography
    (2)
    All Applications
    Filter By Species
    Mouse
    (1)
    Rat
    (3)
    All Species

    Review for MMP-9 (5427-MM) (1) 51

    Average Rating: 5
    (Based on 1 review)

    Reviews using 5427-MM:
    Filter by Applications
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    Images Ratings Applications Species Date Details
     MMP-9 5427-MM
    Enlarge
    reviewed by:
    Anonymous
    04/06/2018
    View

    Summary

    LotSMD0516121
    CommentsThis was a great positive control. It already came in a liquid form so all you had to do was make a dilution factor factor of it in your preferred working buffer. It was very concentrated to begin which allowed us to dilute it to 5ng/ul and 10ng/ul. This will allow us to get a lot out of the product! An example Zymogram showing these to concentrations are attached so you can see how great this works!! This has more uses than one would have imagined.

    Comments

    CommentsThis will be used for a positive control for our future Zymograms and publications.

    FAQs for MMP-9 (5427-MM). (Showing 1 - 1 of 1 FAQs).

    1.  I’m looking for a pair of antibodies to MMP-9 that can be used in a sandwich assay. Do you carry any? 
      • We have 10 primary antibodies for MMP-9 that have been tested in ELISA.https://www.novusbio.com/search?keywords=MMP-9&category=Primary%20Antibodies&applications=ELISA&common_name=MMP-9It looks like 2 have been tested for capture (please note the tested species for each of these).https://www.novusbio.com/search?keywords=MMP-9&category=Primary%20Antibodies&common_name=MMP-9&applications=ELISA%20Capture%20(Matched%20Antibody%20Pair)1 has been tested for detection.https://www.novusbio.com/search?keywords=MMP-9&category=Primary%20Antibodies&common_name=MMP-9&applications=ELISA%20Detection%20(Matched%20Antibody%20Pair)

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    Recent Reviews

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    Anonymous
    04/06/2018
    Application:
    Species:

    Bioinformatics

    Gene Symbol Mmp9
    Uniprot
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