Recombinant Cynomolgus Monkey MMP-9 Protein, CF

Recombinant Cynomologous Monkey MMP-9 Protein (Catalog # 10833-MP) is measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (ES001).

2 μg/lane of Recombinant Cynomolgus Monkey MMP-9 (Catalog # 10833-MP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at ~90 kDa under reducing conditions.

• Reactivity: Pm-Cm
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Cynomolgus Monkey 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 >750 pmol/min/μg, as measured under the described conditions.
• Source: Chinese Hamster Ovary cell line, CHO-derived cynomolgus monkey MMP-9 protein
  Ala20-Asp707
• Accession #: XP_005569271.2
• N-terminal Sequence: Ala20
• Structure / Form: Proform
• Protein/Peptide Type: Recombinant Enzymes
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 76 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: 83-95 kDa, under reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris, CaCl₂, NaCl and Brij-35.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 10 mM CaCl₂, 150 mM NaCl, 0.05% Brij-35 (w/v), pH 7.5 (TCNB)
  • Recombinant Cynomolgus Monkey MMP-9 (rcynoMMP-9) (Catalog # 10833-MP)
  • p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A9563), 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)
  • Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rcynoMMP-9 to 100 µg/mL in Assay Buffer.
  2. Activate rcynoMMP-9 by adding APMA to a final concentration of 1 mM.
  3. Incubate at 37 °C for 24 hours.
  4. Dilute activated rcynoMMP-9 to 0.2 µg/mL in Assay Buffer.
  5. Dilute Substrate to 20 µM in Assay Buffer.
  6. Load 50 µL of 0.2 µg/mL rcynoMMP-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 (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:
  • rcynoMMP-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 Cynomolgus Monkey 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 metalloproteinase 9 (MMP-9), also known as gelatinase B, is a member of the MMP zinc-dependent family of endopeptidases. It cleaves and degrades a variety of targets including important extracellular matrix (ECM) proteins: gelatin, collagen, and elastin, as well as chemokines and extracellular domain plasma membrane proteins (1-3). MMP-9 is synthesized and secreted by several cells including neutrophils, macrophages, fibroblasts, and endothelial cells (4). The monomeric MMP-9 protein is composed of several distinct domains including a signal sequence, a pro-domain which is cleaved upon activation, and a catalytic domain at the n-terminus followed by a hinge region and the c-terminal hemopexin-like domains that contribute to substrate recognition and specificity (5,6). The catalytic domain contains fibronectin type II domains, an active site, and a zinc binding site. MMP-9 can exist as a monomer, disulfide-linked homodimer, or heterodimer in complex with lipocalin‑2 (7,8). MMP-9 activity is regulated at several levels via transcription, post-transcription, translation, secretion, activation, and inhibition. As MMP-9 is involved in ECM remodeling and membrane protein cleavage, it has been widely associated to play a role in several diseases including cancers (9), autoimmune, and cardiovascular diseases (9-11). MMP-9 is consequently an important target of interest for inhibition (11-13). Additionally, it has been found to be a potential biomarker for many types of cancer including pancreatic, osteosarcoma, lung, ovarian, and breast (9).

1. Kridel, S.J. et al. (2001) J. Biol. Chem. 276:20572.
2. Vaisar, T. et al. (2009) Mol. Cell. Proteom. MCP 8:1044.
3. Dufour, A. and C.M. Overall. (2013) Trends Pharmacol. Sci. 34:233.
4. Vandooren, J. et al. (2013) Crit. Rev. Biochem. Mol. Biol. 48:222.
5. Roeb, E. et al. (2002) J. Biol. Chem. 277:50326.
6. Rosenblum, G. et al. (2007) Structure 15:1227.
7. Kjeldsen, L. et al. (1993) J. Biol. Chem. 268:10425.
8. Olson, M.W. et al. (2000) J. Biol. Chem. 275:2661.
9. Huang, H. (2018) Sensors 18:3249.
10. Ram, M. et al. (2006) J. Clin. Immunol. 26:299.
11. Hu, J. et al. (2007) Nat. Rev. Drug. Discov. 6:480.
12. Fields, G.B. (2019) Cells. 8:984.
13. Kumar, G.B. et al. (2019) Medchemcomm. 10:2024.

FAQs for MMP-9 (10833-MP). (Showing 1 - 1 of 1 FAQ).

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, seen here.It looks like 2 have been tested for capture (please note the tested species for each of these), seen here.6 have been tested for detection, seen here.

Bioinformatics

• Uniprot:
  • XP_005569271.2()