Recombinant Human MMP-12 Protein, CF

Newer Version Available: 917-MPB
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Discontinued Product

917-MP has been discontinued and is replaced by 917-MPB.

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Recombinant Human MMP-12 Protein, CF Summary

Product Specifications

Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001). The specific activity is >500 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human MMP-12 protein
Leu17-Cys470
Accession #
N-terminal Sequence
Analysis
Leu17
Structure / Form
Pro form
Predicted Molecular Mass
52 kDa
SDS-PAGE
56 kDa, reducing conditions

Product Datasheets

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917-MP

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

917-MP

Formulation Supplied as a 0.2 μm filtered solution in MES, 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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Human MMP-12 (rhMMP-12) (Catalog # 917-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
  1. Dilute rhMMP-12 to 50 µg/mL in Assay Buffer.
  2. Activate rhMMP-12 by adding APMA to a final concentration of 1 mM.
  3. Incubate at 37 °C for 4 hours to activate.
  4. Dilute activated rhMMP-12 to 0.4 ng/µL in Assay Buffer.
  5. Dilute Substrate to 20 µM in Assay Buffer.
  6. Load into plate 50 µL of 0.4 ng/µL rhMMP-12, 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.
  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:
  • rhMMP-12: 0.020 µg
  • Substrate: 10 µM
Reconstitution Calculator

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Background: MMP-12

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-12 (macrophage elastase) is found in macrophages and its expression in monocytes can be induced by cytokines such as GM-CSF and CD40 signaling (1). In addition to elastin, MMP-12 can degrade a broad spectrum of substrates, including type IV collagen, fibronectin, laminin, vitronectin, proteoglycans, chondroitin sulfate, myelin basic protein, alpha 1-antitrypsin, and plasminogen. It can also activate MMP-2 and MMP-3. MMP-12 is required for macrophage-mediated proteolysis and matrix invasion in mice. MMP-12 is proposed to have a direct role in the pathogenesis of aortic aneurysms and in the development of pulmonary emphysema that results from chronic inhalation of cigarette smoke. Structurally, the pro MMP-12 consists of following domains: a pro domain, a catalytic domain containing the zinc-binding site, and a C-terminal hemopexin-like domain. The rhMMP-12 corresponds to the pro form that can be activated by autocatalysis under the conditions described above.

References
  1. S.D. Shapiro et al. (2004) in Handbook of Proteolytic Enzymes (eds. A.J. Barrett et al.) pp.540 - 544, Academic Press, San Diego.
Long Name
Matrix Metalloproteinase 12
Entrez Gene IDs
4321 (Human); 17381 (Mouse)
Alternate Names
EC 3.4.24; hME; HMEEC 3.4.24.65; Macrophage elastase; macrophage metalloelastase; matrix metallopeptidase 12 (macrophage elastase); matrix metalloproteinase 12 (macrophage elastase); Matrix metalloproteinase-12; ME; MGC138506; MME; MMP12; MMP-12

Citations for Recombinant Human MMP-12 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.

12 Citations: Showing 1 - 10
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  1. 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
  2. Fc-engineered antibodies with immune effector functions completely abolished
    Authors: I Wilkinson, S Anderson, J Fry, LA Julien, D Neville, O Qureshi, G Watts, G Hale
    PLoS ONE, 2021-12-21;16(12):e0260954.
    Species: Human
    Sample Types: Recombinant Proteins
    Applications: Bioassay
  3. 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
  4. MMP-12, Secreted by Pro-Inflammatory Macrophages, Targets Endoglin in Human Macrophages and Endothelial Cells
    Authors: M Aristorena, E Gallardo-V, M Vicen, M de Las Cas, L Ojeda-Fern, C Nieto, FJ Blanco, AC Valbuena-D, LM Botella, P Nachtigal, AL Corbi, M Colmenares, C Bernabeu
    Int J Mol Sci, 2019-06-25;20(12):.
    Species: Human
    Sample Types: Small Molecule
    Applications: Activation
  5. 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
  6. Development of matrix metalloproteinase-targeted probes for lung inflammation detection with positron emission tomography
    Authors: N Kondo, T Temma, K Aita, S Shimochi, K Koshino, M Senda, H Iida
    Sci Rep, 2018-01-22;8(1):1347.
    Applications: Bioassay
  7. Characterizing the O-glycosylation landscape of human plasma, platelets, and endothelial cells
    Authors: SL King, HJ Joshi, KT Schjoldage, A Halim, TD Madsen, MH Dziegiel, A Woetmann, SY Vakhrushev, HH Wandall
    Blood Adv, 2017-02-23;1(7):429-442.
    Species: Human
    Sample Types:
    Applications: Bioassay
  8. Serological assessment of neutrophil elastase activity on elastin during lung ECM remodeling.
    Authors: Kristensen J, Karsdal M, Sand J, Willumsen N, Diefenbach C, Svensson B, Hagglund P, Oersnes-Leeming D
    BMC Pulm Med, 2015-05-03;15(0):53.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  9. MMP mediated degradation of type IV collagen alpha 1 and alpha 3 chains reflects basement membrane remodeling in experimental and clinical fibrosis--validation of two novel biomarker assays.
    Authors: Sand J, Larsen L, Hogaboam C, Martinez F, Han M, Rossel Larsen M, Nawrocki A, Zheng Q, Karsdal M, Leeming D
    PLoS ONE, 2013-12-23;8(12):e84934.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  10. 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
  11. Regulation of progranulin expression in human microglia and proteolysis of progranulin by matrix metalloproteinase-12 (MMP-12).
    Authors: Suh HS, Choi N, Tarassishin L, Lee SC
    PLoS ONE, 2012-04-11;7(4):e35115.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  12. 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

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