Recombinant Mouse MMP-8 Protein, CF

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Product Details

Summary
Reactivity MuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Mouse MMP-8 Protein, CF Summary

Details of Functionality
Measured by its ability to cleave a fluorogenic peptide substrate Mca-KPLGL-Dpa-AR-NH2 (Catalog # ES010). The specific activity is >400 pmol/min/µg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived mouse MMP-8 protein
Phe21-Ser465, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Phe21 & Leu27
Structure / Form
Pro form
Protein/Peptide Type
Recombinant Enzymes
Gene
Mmp8
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
52 kDa and 51 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
60-65 kDa doublet, reducing conditions
Publications
Read Publications using
2904-MP 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, -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 MES, NaCl, CaCl2 and Brij-35.
Purity
>95%, 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% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Mouse MMP-8 (rmMMP-8) (Catalog # 2904-MP)
  • p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A-9563), 100 mM stock in DMSO
  • Substrate MCA-Lys-Pro-Leu-Gly-Leu-DAP(DNP)-Ala-Arg-NH2 (Catalog # ES010), 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 rmMMP-8 to 20 µg/mL in Assay Buffer.
  2. Activate rmMMP-8 by adding APMA to a final concentration of 1 mM.
  3. Incubate at room temperature for 2 hours.
  4. Dilute activated rmMMP-8 to 0.5 ng/µL in Assay Buffer.
  5. Dilute Substrate to 80 µM in Assay Buffer.
  6. Load into a black well plate 50 µL of 0.5 ng/µL rmMMP-8, and start the reaction by adding 50 µL of 80 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 80 µM Substrate without any rmMMP-8.
  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:
  • rmMMP-8: 0.025 μg
  • Substrate: 40 µM

Notes

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

Alternate Names for Recombinant Mouse MMP-8 Protein, CF

  • CLG1HNC
  • Collagenase 2
  • EC 3.4.24
  • EC 3.4.24.34
  • matrix metallopeptidase 8 (neutrophil collagenase)
  • matrix metalloproteinase 8 (neutrophil collagenase)
  • Matrix metalloproteinase-8
  • MMP8
  • MMP-8
  • neutrophil collagenase
  • PMNL collagenase
  • PMNL-CL

Background

Matrix metalloproteinases (MMPs) are a family of zinc and calcium dependent endopeptidases with the combined ability to degrade many components of the extracellular matrix. MMP-8 (neutrophil collagenase) is expressed in neutrophils, where it is stored in specific granules. MMP-8 release from the neutrophils is stimulated by various factors such as interleukins 1 and 8, TNF-alpha and GM-CSF. MMP-8 is capable of cleaving types I, II and III triple-helical collagen, gelatin peptides, fibronectin, proteoglycans, aggrecan, serpins, beta -casein and peptides such as angiotensin and substance P. In addition to its function in phagocytosis,
MMP‑8 has a high capacity for infiltrating connective tissue, and is implicated in the breakdown of the extracellular matrix in diseases such as rheumatoid arthritis. Structurally, MMP-8 consists of several domains: a pro-domain that is cleaved upon activation, a catalytic domain containing the zinc-binding site, a short hinge region and a hemopexin-like domain.

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Publications for MMP-8 (2904-MP)(4)

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Bioinformatics

Gene Symbol Mmp8
Entrez
Uniprot