Recombinant Human MMP-13 Protein, CF Summary
Product Specifications
Leu20-Cys471
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.
511-MM
| Formulation | Supplied as a 0.2 μm filtered solution in MES, NaCl, CaCl2 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.
|
Assay Procedure
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35 (w/v), pH 7.5 (TCNB)
- Recombinant Human MMP-13 (rhMMP-13) (Catalog # 511-MM)
- p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563) 100 mM stock in DMSO
- Fluorogenic Peptide Substrate I: MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Catalog # ES001)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhMMP-13 to 100 µg/mL in Assay Buffer.
- Add APMA to a final concentration of 1 mM.
- Incubate at 37 °C for 2 hours to activate.
- Dilute activated rhMMP-13 to 0.2 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of the 0.2 ng/µL rhMMP-13 into a black well plate and start the reaction by adding 50 µL of 20 µM Substrate. 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 (top read), 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-13: 0.010 µg
- Substrate: 10 µM
Reconstitution Calculator
Background: MMP-13
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-13 (Collagenase-3) has been demonstrated to degrade a range of extracellular matrix proteins, including collagen types I, II, III, IV, IX, X and XIV, gelatin, aggrecan, perlecan and fibronectin. MMP-13 is distinguished from the other human collagenases by its effecient degradation of type II collagen. MMP-13 is expressed by fibroblasts, chrondrocytes and squamous epithelial cells. Structurally, MMP-13 may be divided into several distinct domains; a pro-domain which is cleaved upon activation; a catalytic domain containing the zinc binding site; a short hinge region and a carboxyl terminal (hemopexin-like) domain.
- Jeffery, J.J. (1998) in Collagenase 3. A.J. Barrett, et al. (eds): Handbook of Proteolytic Enzymes, San Diego: Academic Press, p. 1167.
Citations for Recombinant Human MMP-13 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.
11
Citations: Showing 1 - 10
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MMP13-targeted siRNA-loaded micelles for diagnosis and treatment of posttraumatic osteoarthritis
Authors: Zhou, D;Wei, Y;Sheng, S;Wang, M;Lv, J;Zhao, B;Chen, X;Xu, K;Bai, L;Wu, Y;Song, P;Cao, L;Zhou, F;Zhang, H;Shi, Z;Su, J;
Bioactive materials
Species: N/A
Sample Types: RNA
Applications: Bioassay -
dsRNA Induced IFNbeta-MMP13 Axis Drives Corneal Wound Healing
Authors: X Lan, W Zhang, J Zhu, H Huang, K Mo, H Guo, L Zhu, J Liu, M Li, L Wang, C Liu, J Ji, H Ouyang
Investigative Ophthalmology & Visual Science, 2022-02-01;63(2):14.
Species: Mouse
Sample Types: Whole Cells
Applications: Tissue Culture -
Crenigacestat, a selective NOTCH1 inhibitor, reduces intrahepatic cholangiocarcinoma progression by blocking VEGFA/DLL4/MMP13 axis
Authors: S Mancarella, G Serino, F Dituri, A Cigliano, S Ribback, J Wang, X Chen, DF Calvisi, G Giannelli
Cell Death Differ., 2020-02-10;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: Cell Culture -
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 -
Development of an inflammatory tissue-selective chimeric TNF receptor
Authors: CJ Lee, CC Wang, M Chen, KH Chuang, TL Cheng, TY Jian, YM Wang, TH Huang, KW Liao, SC Tzou
Cytokine, 2018-10-21;0(0):.
Species: Mouse
Sample Types: Recombinant Protein
Applications: Bioassay -
Novel Arginine-containing Macrocyclic MMP Inhibitors: Synthesis, 99mTc-labeling, and Evaluation
Authors: Y Ye, J Toczek, K Gona, HY Kim, J Han, M Razavian, R Golestani, J Zhang, TL Wu, M Ghosh, JJ Jung, MM Sadeghi
Sci Rep, 2018-08-03;8(1):11647.
Species: Human
Sample Types: Peptide
Applications: Enzyme Assay -
The effect of protease inhibitors on the induction of osteoarthritis-related biomarkers in bovine full-depth cartilage explants.
Authors: He Y, Zheng Q, Jiang M, Sun S, Christiansen T, Kassem M, Karsdal M, Bay-Jensen A
PLoS ONE, 2015-04-24;10(0):.
Applications: Western Blot -
IL-1beta-induced matrix metalloproteinase-13 is activated by a disintegrin and metalloprotease-28-regulated proliferation of human osteoblast-like cells.
Authors: Ozeki N, Kawai R, Yamaguchi H, Hiyama T, Kinoshita K, Hase N, Nakata K, Kondo A, Mogi M, Nakamura H
Exp Cell Res, 2014-03-05;323(1):165-77.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Resistance of corneal RFUVA-cross-linked collagens and small leucine-rich proteoglycans to degradation by matrix metalloproteinases.
Authors: Zhang Y, Mao X, Schwend T, Littlechild S, Conrad G
Invest Ophthalmol Vis Sci, 2013-02-05;54(2):1014-25.
Species: Bovine
Sample Types: 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 -
Chondrocyte hypertrophy and apoptosis induced by GROalpha require three-dimensional interaction with the extracellular matrix and a co-receptor role of chondroitin sulfate and are associated with the mitochondrial splicing variant of cathepsin B.
Authors: Olivotto E, Vitellozzi R, Fernandez P, Falcieri E, Battistelli M, Burattini S, Flamigni F, Santi S, Borzi RM
J. Cell. Physiol., 2007-02-01;210(2):417-27.
Applications: ELISA (Standard)
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-13 Protein, CF
Average Rating: 5 (Based on 2 Reviews)
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Reason for Rating: The protein was very practical to dissolve, aliquot, and use without losing enzymatic activity.
rhMMP-13 was used as a control to induce inflammation in human THP-1 monocytes differentiated into mature macrophages. Monocytes were treated with lipopolysaccharide (LPS, 1 microgram/mL) and rhMMP-13 (10-100 ng/mL). Results showed a dose-dependent generation of nitrites in these cells, confirming the pro-inflammatory activity of rhMMP-13.
An ELISA was developed to measure MMP-13 in human plasma samples. This recombinant protein was used in this assay to confirm functionality and to build a standard curve.
