Home / MMP-12 / Recombinant Human MMP-12 Protein, CF

Recombinant Human MMP-12 Protein, CF

Catalog #: 917-MPB
12 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
917-MPB-020
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Recombinant Human MMP-12 Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 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, with Val97Leu substitution
Accession #
P39900
N-terminal Sequence
Analysis
Leu17
Structure / Form
Pro form
Predicted Molecular Mass
52 kDa
SDS-PAGE
53-60 kDa, reducing conditions

Product Datasheets

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917-MPB

Product Datasheet
COA
SDS

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-MPB

Formulation Lyophilized from a 0.2 μm filtered solution in MES, NaCl, CaCl2, CHAPS, ZnSO4 and PEG.
Reconstitution Reconstitute at 0.25 mg/mL in 10 mM MES, 0.1 M NaCl, 100 μM CaCl2, 0.1% CHAPS, 1 μM ZnSO4 and 0.1% PEG., pH 6.0.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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-MPB)
  • p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A9563), 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 and incubating at 37 °C for 24 hours.
  3. Dilute activated rhMMP-12 to 0.4 ng/µL in Assay Buffer.
  4. Dilute Substrate to 20 µM in Assay Buffer.
  5. Load into plate 50 µL of 0.4 ng/µL rhMMP-12, and start the reaction by adding 50 µL of 20 µM Substrate.
  6. 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.
    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

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: MMP-12

Matrix metalloproteinases (MMPs) are a family of zinc and calcium dependent endopeptidases that collectively degrade the components of the extracellular matrix. MMP-12 (macrophage elastase) consists of the following domains: a pro domain, a catalytic domain containing the zinc-binding site, and a C-terminal hemopexin-like domain. The 54 kDa rhMMP-12 pro form is activated via processing into 45 kDa and 22 kDa active forms (1). MMP-12 is capable of cleaving several substrates in addition to elastin. Macrophage secretion of MMP-12 at sites of inflammation can be induced by cytokines. Given the secretion of MMP12 during an inflammatory response, MMP-12 is involved in many pathologies including vascular disease (2, 3) and cancer (4-6). In particular, MMP12-mediated pathological degradation of the extracellular matrix is a well-established key event in inflammatory-related pulmonary disease (7). For example, overexpression of MMP12 in alveolar macrophages is associated with smoking and emphysema (8) while different MMP-12 variants result in protection against chronic obstructive pulmonary disease (COPD) development (9) or cause increased risk and disease severity (10). MMP-12 has also been shown to translocate into the nucleus of viral-infected cells to directly regulate transcription during an immune response (11).

References
  1. Shapiro, S.D. et al. (1993) J. Biol. Chem. 268:23824.
  2. Liu, S.L. et al. (2015) Sci. Rep. 5:17189.
  3. Iyer, R.P. et al. (2015) Int. J. Cardiol. 185:198.
  4. Cao, W. et al. (2017) Oncol. Rep. 3:1401.
  5. Klupp, F. et al. (2016) BMC Cancer 16:494.
  6. Chung, I.C. et al. (2014) BMC Cancer 14:348.
  7. Vanderbroucke, R.E. et al. (2011) Eur. Respir. J. 387:1200.
  8. Babusyte, A. et al. (2007) Respir. Res. 8:81.
  9. Hunninghake, G.M. et al. (2009) N. Engl. J. Med. 361:2599.
  10. Mukhopadhyay, S. et al. (2010) J. Allergy Clin. Immunol. 126:70.
  11. Marchant, D.J. et al. (2013) Nature Medicine 20:493.
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

FAQs

  1. 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.

  2. 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.

  3. Does this MMP enzyme need to be activated to work?

    • Yes, this enzyme requires activation prior to use.

  4. 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.

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