Recombinant Human MMP-9 His-tag Avi-tag Protein, CF

R&D Systems | Catalog # AVI911

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human MMP-9 His-tag Avi-tag Protein (AVI911)
  • 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

Structure / Form

Proform

Conjugate

Biotin

Applications

Binding Activity, Enzyme Activity
View all MMP-9 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human MMP-9 protein
Human MMP-9
(Ala20-Asp707)
Accession # P14780.1		 6-His tag GG Avi-tag
N-terminus C-terminus

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ala20

Predicted Molecular Mass

79.0 kDa

SDS-PAGE

90-100 kDa, under reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001).
The specific activity is >1500 pmol/min/μg, as measured under the described conditions.

Measured by its binding ability in a functional ELISA.
Biotinylated Recombinant Human MMP-9 His-tag Avi-tag (Catalog # AVI911) binds Human/Primate MMP-9 Antibody (Catalog # MAB936R) with an ED50 of 1.25-15.0 ng/mL.

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

Biotinylated Recombinant Human MMP‑9 His-tag Avi-tag Protein Enzyme Activity.

Recombinant Human MMP-9 His-tag Avi-tag (Catalog # AVI911) is measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (ES001).

Biotinylated Recombinant Human MMP‑9 His-tag Avi-tag Protein Binding Activity.

Biotinylated Recombinant Human MMP-9 His-tag Avi-tag Protein (Catalog # AVI911) binds Human/Primate MMP-9 Antibody (MAB936R) with an ED50 of 1.25-15.0 ng/mL.

Biotinylated Recombinant Human MMP‑9 His-tag Avi-tag Protein SDS-PAGE.

2 μg/lane of Biotinylated Recombinant Human MMP‑9 His-tag Avi-tag Protein (Catalog # AVI911) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 90-100 kDa under reducing conditions.

Formulation, Preparation, and Storage

AVI911
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

Recombinant human Active Matrix Metalloproteinase 9 (MMP-9), also known as gelatinase B, is a member of the MMP family of zinc and calcium-dependent endopeptidases. MMP-9 protein is synthesized as a pre-proenzyme in specific cells such as neutrophils, macrophages, fibroblasts and endothelial cells (1). Expressed MMP-9 contains a signal peptide to transport it to the extracellular matrix (ECM), a hinge region, a propeptide region, a catalytic domain, and a hemopexin-like domain that is important for substrate recognition (2,3).  In addition to having three fibronectin type II domains that contribute to substrate binding and an active site, the catalytic domain of MMP-9 also contains a zinc-binding region that interacts with a cysteine in the propeptide to maintain latency.  Consequently, removal of the propeptide through cleavage by proteases in the ECM, such as MMP3, activates the protein (2, 3). MMP-9 has specificity for targets containing an established preferred consensus sequence (3-5) and has a broad range of substrates within the ECM including gelatin, collagen, elastin that contributes to its role in ECM remodeling and extracellular domain cell surface protein release from the plasma membrane (3). Due to its activity in the ECM, MMP-9 plays a role in many biological processes and can serve as a biomarker in tumor invasion and mestastasis (3,6) of many cancers including colon, ovarian, breast, osteosarcoma, and lung cancers (7-11) making it a therapeutic target (6, 8, 12). MMP-9 modeling of the ECM also leads to a pivotal role in other inflammation- and autoimmune-related diseases (3, 13, 14). Our Avi-tag Biotinylated human MMP-9 His-tag features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

References

  1. Vandooren, J. et al. (2013) Crit. Rev. Biochem. Mol. Biol.  48: 222.
  2. Roeb, E. et al. (2002) J. Biol. Chem. 277: 50326.
  3. Huang, H. (2018) Sensors. 18:3249.
  4. Kridel, S.J. et al. (2001) J. Biol. Chem. 276: 20572.
  5. Prudova, A. et al. (2010) Mol. Cell Proteom. 9:894.
  6. Kalali, D. (2023) Glob. Med. Genet. 10:48.
  7. Hu, X. et al. (2012) Arch. Glynecol. Obstet. 286:1537.
  8. Wang, J. et al. (2014) Clin. Chim. Acta. 433:225.
  9. Blanco-Prieto, S. et al. (2017) BMC Cancer 17:823.
  10. Liang, S. and L. Chang. (2018) Biomark. Med. 12:393.
  11. Malik, S. et al. (2024) Sci. Rep. 14:15117.
  12. Roy, R. et al. (2009) J. Clin. Oncol. 27:5287.
  13. Ram, M. et al. (2006) J. Clin. Immunol. 26:299.
  14. Kim, I.S. et al. (2023) Curr. Med. Chem. 30:2075.

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

Additional MMP-9 Products

Product Documents for Recombinant Human MMP-9 His-tag Avi-tag Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human MMP-9 His-tag Avi-tag Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human MMP-9 His-tag Avi-tag Protein, CF (AVI911):

Materials
  • Assay Buffer: 50 mM Tris, 150 mM NaCl, 10 mM CaCl2, 0.05% Brij-35 (w/v), pH 7.5 (TCNB)
  • Recombinant Human MMP-9 His-tag Avi-tag (rhMMP-9) (Catalog # AVI911)
  • p-aminophenylmercuric acetate (APMA), 100 mM stock in DMSO 
  • Substrate: MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES001)
  • Black 96-well Plate  
  • Plate Reader with Fluorescence Read Capability 
  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 µg/mL in Assay Buffer. 
  5. Dilute Substrate to 40 µM in Assay Buffer. 
  6. Load 50 µL of 0.4 µg/mL rhMMP-9 into a plate and start the reaction by adding 50 µL of 40 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 40 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
Per Well:
rhMMP-9: 0.02 µg
Substrate: 20 µM 

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