Recombinant Human MMP-8 Protein, CF Summary
Product Specifications
Phe21-Gly467
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
908-MP
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Assay Procedure
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Recombinant Human MMP‑8 (rhMMP-8) (Catalog # 908-MP)
- 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
- Activate rhMMP-8 at 100 µg/mL with 1 mM APMA in Assay Buffer.
- Incubate reaction at 37 °C for 1 hour.
- Dilute activated rhMMP-8 to 1.0 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- In a plate load 50 µL of 1.0 ng/µL rhMMP-8, and start the reaction by adding 50 µL of 20 µM Substrate to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
- Read at excitation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes.
- 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-8: 0.050 µg
- Substrate: 10 µM
Reconstitution Calculator
Background: MMP-8
Matrix metalloproteinases (MMPs) are a family of zinc and calcium dependent endopeptidases with the combined ability to degrade all the 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. MMP-8 is heavily glycosylated.
Citations for Recombinant Human MMP-8 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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A molecular interactome of the glioblastoma perivascular niche reveals integrin binding sialoprotein as a mediator of tumor cell migration
Authors: Y Ghochani, SD Muthukrish, A Sohrabi, R Kawaguchi, MC Condro, S Bastola, F Gao, Y Qin, J Mottahedeh, ML Iruela-Ari, N Rao, DR Laks, LM Liau, GW Mathern, SA Goldman, ST Carmichael, I Nakano, G Coppola, SK Seidlits, HI Kornblum
Cell Reports, 2022-10-18;41(3):111511.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Loss of mutual protection between human osteoclasts and chondrocytes in damaged joints initiates osteoclast-mediated cartilage degradation by MMPs
Authors: QC Larrouture, AP Cribbs, SR Rao, M Philpott, SJ Snelling, HJ Knowles
Scientific Reports, 2021-11-22;11(1):22708.
Species: Human
Sample Types: Recombinant Proteins
Applications: Zymography Control -
MMP8 increases tongue carcinoma cell-cell adhesion and diminishes migration via cleavage of anti-adhesive FXYD5
Authors: K Juurikka, A Dufour, K Pehkonen, B Mainoli, P Campioni R, N Solis, T Klein, P Nyberg, CM Overall, T Salo, P Åström
Oncogenesis, 2021-05-31;10(5):44.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Concerted actions by MMPs, ADAMTS and serine proteases during remodeling of the cartilage callus into bone during osseointegration of hip implants
Authors: J Cassuto, A Folestad, J Göthlin, H Malchau, J Kärrholm
Bone Rep, 2020-09-11;13(0):100715.
Species: Human
Sample Types: Plasma
Applications: ELISA Detection -
Matrix Metalloproteinase Triple-Helical Peptide Inhibitors: Potential Cross-Reactivity with Caspase-11
Authors: AM Knapinska, M Hart, G Drotleff, GB Fields
Molecules, 2019-11-28;24(23):.
Species: Human
Sample Types: Peptide
Applications: Bioassay -
Breast cancer cells rely on environmental pyruvate to shape the metastatic niche
Authors: I Elia, M Rossi, S Stegen, D Broekaert, G Doglioni, M van Gorsel, R Boon, C Escalona-N, S Torrekens, C Verfaillie, E Verbeken, G Carmeliet, SM Fendt
Nature, 2019-02-27;0(0):.
Species: Human
Sample Types: Spheroids
Applications: Bioassay -
Lytic and mechanical stability of clots composed of fibrin and blood vessel wall components.
Authors: Rottenberger Z, Komorowicz E, Szabo L, Bota A, Varga Z, Machovich R, Longstaff C, Kolev K
J Thromb Haemost, 2013-03-01;11(3):529-38.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay -
Simple pseudo-dipeptides with a P2' glutamate: a novel inhibitor family of matrix metalloproteases and other metzincins.
Authors: Devel L, Beau F, Amoura M, Vera L, Cassar-Lajeunesse E, Garcia S, Czarny B, Stura E, Dive V
J Biol Chem, 2012-06-11;287(32):26647-56.
Applications: Enzyme Assay -
Astacin proteases cleave dentin sialophosphoprotein (Dspp) to generate dentin phosphoprotein (Dpp).
Authors: Tsuchiya S, Simmer JP, Hu JC, Richardson AS, Yamakoshi F, Yamakoshi Y
J. Bone Miner. Res., 2011-01-01;26(0):220.
Species: Human
Sample Types:
Applications: Bioassay -
Matrix metalloprotease-9 dysregulation in lower airway secretions of cystic fibrosis patients.
Authors: Gaggar A, Li Y, Weathington N, Winkler M, Kong M, Jackson P, Blalock JE, Clancy JP
Am. J. Physiol. Lung Cell Mol. Physiol., 2007-03-23;293(1):L96-L104.
Applications: Western Blot
FAQs
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Can the enzyme be stored after activation, or do I need to use it immediately after activation?
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.
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If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?
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.
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Does this MMP enzyme need to be activated to work?
Yes, this enzyme requires activation prior to use.
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What is the activity of this enzyme in units/µg?
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.
Reviews for Recombinant Human MMP-8 Protein, CF
Average Rating: 5 (Based on 3 Reviews)
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Reason for Rating: The protein did not lose any enzymatic activity after storing and freezing, was easy to dissolve and allowed us to have replicable results in vitro
rhMMP-8 was used to induce pro-inflammatory responses in ARPE-19 human cells. After the induction, the present of pro-angiogenic proteins, a proteolytic product derived from the pro-inflammatory status, was quantified by confocal imaging.
