Recombinant Human MMP-3 Protein, CF Summary
Product Specifications
Tyr18-Cys477 (Lys45Glu)
Analysis
Product Datasheets
513-MP
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.
513-MP
| Formulation | Supplied as a 0.2 μm filtered solution in MES, NaCl, CaCl2, Glycerol 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.
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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-3 (rhMMP-3) (Catalog # 513-MP)
- Chymotrypsin (Sigma, Catalog # C-3142), 1 mg/mL stock in 1 mM HCl
- Phenylmethyl Sulfonyl Fluoride (PMSF) (Sigma, Catalog # P-7626), 0.2 M stock in 2-Propanol
- Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002), 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-3 at 20 µg/mL in Assay Buffer containing 5 µg/mL Chymotrypsin.
- Incubate reaction at 37 °C for 30 minutes.
- Stop activation with 2 mM PMSF. Pre-warm the PMSF to 37 °C prior to adding to sample.
- Dilute activated rhMMP-3 to 2.5 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- In a plate load 50 µL of 2.5 ng/µL rhMMP-3, 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 (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).
- rhMMP-3: 0.125 µg
- Substrate: 10 µM
Background: MMP-3
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-3 (stromelysin-1), can degrade a broad range of substrates including collagen alpha chains, aggrecan, laminin, fibronectin, elastin, casein, alpha -1 antitrypsin, myelin basic protein, IL-1 beta, IGFBP-3, pro MMP-1, pro MMP-7, pro MMP-8, pro MMP-9 and pro MMP-13. MMP-3 does not cleave the triple helical region of interstitial collagens, a characteristic which distinguishes the stromelysins from the collagenases. The MMP-3 substrate repertoire extends beyond extracellular matrix proteins and implicates MMP-3 in roles other than direct tissue remodelling, for instance, enzyme cascades and cytokine regulation. MMP-3 is expressed by fibroblasts, chrondrocytes, osteoblasts, endothelial cells, smooth muscle cells and macrophages. Structurally, MMP-3 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.
Citations for Recombinant Human MMP-3 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.
9
Citations: Showing 1 - 9
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Preclinical development of a bispecific TNFalpha/IL-23 neutralising domain antibody as a novel oral treatment for inflammatory bowel disease
Authors: KJ Roberts, MF Cubitt, TM Carlton, L Rodrigues-, L Maggiore, R Chai, S Clare, K Harcourt, TT MacDonald, KP Ray, A Vossenkämp, MR West, JS Crowe
Scientific Reports, 2021-09-30;11(1):19422.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Regional contribution of proteoglycans to the fracture toughness of the dentin extracellular matrix
Authors: Y Alania, J Creighton, LT Trevelin, CA Zamperini, AK Bedran-Rus
J Biomech, 2020-01-16;0(0):109633.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
Biodistribution of Nanostructured Lipid Carriers in Mice Atherosclerotic Model
Authors: L Devel, G Almer, C Cabella, F Beau, M Bernes, P Oliva, F Navarro, R Prassl, H Mangge, I Texier
Molecules, 2019-09-26;24(19):.
Species: Human
Sample Types: Peptide
Applications: Bioassay -
Preclinical Development of a Novel, Orally-Administered Anti-Tumour Necrosis Factor Domain Antibody for the Treatment of Inflammatory Bowel Disease
Authors: JS Crowe, KJ Roberts, TM Carlton, L Maggiore, MF Cubitt, S Clare, K Harcourt, J Reckless, TT MacDonald, KP Ray, A Vossenkämp, MR West
Sci Rep, 2018-03-21;8(1):4941.
Applications: Bioassay -
Senescent peritoneal mesothelium creates a niche for ovarian cancer metastases
Cell Death Dis, 2016-12-29;7(12):e2565.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Fibulin-3, -4, and -5 are highly susceptible to proteolysis, interact with cells and heparin, and form multimers.
Authors: Djokic J, Fagotto-Kaufmann C, Bartels R, Nelea V, Reinhardt D
J Biol Chem, 2013-06-19;288(31):22821-35.
Species: Human
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 -
Pre-analytical effects of blood sampling and handling in quantitative immunoassays for rheumatoid arthritis.
Authors: Zhao X, Qureshi F, Eastman PS, Manning WC, Alexander C, Robinson WH, Hesterberg LK
J. Immunol. Methods, 2012-02-17;378(1):72-80.
Applications: ELISA (Standard) -
Targeting ADAM-mediated ligand cleavage to inhibit HER3 and EGFR pathways in non-small cell lung cancer.
Authors: Zhou BB, Peyton M, He B, Liu C, Girard L, Caudler E, Lo Y, Baribaud F, Mikami I, Reguart N, Yang G, Li Y, Yao W, Vaddi K, Gazdar AF, Friedman SM, Jablons DM, Newton RC, Fridman JS, Minna JD, Scherle PA
Cancer Cell, 2006-07-01;10(1):39-50.
Species: Human
Sample Types:
Applications: Enzyme Assay
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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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-3 Protein, CF
Average Rating: 5 (Based on 3 Reviews)
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Reason for Rating: Protein demonstrated a strong pro-inflammatory effect in vivo without losing its enzymatic activity after dissolving, aliquoting, and freezing.
