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Recombinant Mouse/Rat MMP-2 Protein, CF

Catalog #: 924-MP
19 Citations Datasheet / COA / SDS
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924-MP-010
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Citations (19)
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Recombinant Mouse/Rat MMP-2 Protein, CF Summary

Product Specifications

Purity
>95%, 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 >1,500 pmol/min/μg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived MMP-2 protein
Ile34-Cys662
Accession #
P33434
N-terminal Sequence
Analysis
Ile34
Structure / Form
Pro form
Predicted Molecular Mass
71 kDa
SDS-PAGE
72 kDa, under reducing conditions

Product Datasheets

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924-MP

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.

924-MP

Formulation Supplied as a 0.2 μm filtered solution in Tris, CaCl2, NaCl, Brij-35 and Glycerol.
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 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 Mouse/Rat MMP-2 (rm/rMMP-2) (Catalog # 924-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)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rm/rMMP-2 to 40 µg/mL with 1 mM APMA in Assay Buffer.
  2. Incubate at 37 °C for 2 hours to activate rm/rMMP-2.
  3. Dilute activated rm/rMMP-2 to 0.2 ng/µL in Assay Buffer.
  4. Dilute Substrate to 80 µM in Assay Buffer.
  5. Load 50 µL of 0.2 ng/µL rm/rMMP-2 in a plate, and start the reaction by adding 50 µL of 80 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 80 µM Substrate.
  6. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. 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:
  • rm/rMMP-2: 0.010 µg
  • Substrate: 40 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-2

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-2 (gelatinase A), a type IV collagenase, can degrade a broad range of substrates including type IV, V, VII and X collagens as well as elastin and fibronectin. It is believed to act synergistically with interstitial collagenase (MMP-1) in the degradation of fibrillar collagens as it degrades their denatured gelatin forms. MMP-2 has been shown to be associated with many connective tissue cells as well as neutrophils, macrophages and monocytes. Structurally, MMP-2 may be divided into several distinct domains: a pro-domain which is cleaved upon activation; a catalytic domain containing the zinc binding site; a fibronectin-like domain thought to play a role in substrate targeting; and a carboxyl terminal (hemopexin-like) domain containing 2 N-linked glycosylation sites. The amino acid sequences of the proenzymes are identical between mouse and rat.  

Long Name
Matrix Metalloproteinase 2
Entrez Gene IDs
4313 (Human); 17390 (Mouse)
Alternate Names
72 kDa gelatinase; CLG4; CLG4A72 kDa type IV collagenase; collagenase type IV-A; EC 3.4.24; EC 3.4.24.24; Gelatinase A; matrix metallopeptidase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IVcollagenase); matrix metalloproteinase 2 (gelatinase A, 72kD gelatinase, 72kD type IVcollagenase); Matrix metalloproteinase-2; matrix metalloproteinase-II; MMP2; MMP-2; MMP-II; MONA; neutrophil gelatinase; TBE-1matrix metalloproteinase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IVcollagenase)

Citations for Recombinant Mouse/Rat MMP-2 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.

19 Citations: Showing 1 - 10
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  1. Effects of Pera Orange Juice and Moro Orange Juice in Healthy Rats: A Metabolomic Approach
    Authors: Fujimori, ASS;Ribeiro, APD;Pereira, AG;Dias-Audibert, FL;Tonon, CR;Dos Santos, PP;Dantas, D;Zanati, SG;Catharino, RR;Zornoff, LAM;Azevedo, PS;de Paiva, SAR;Okoshi, MP;Lima, EO;Polegato, BF;
    Metabolites
    Species: Rat
    Sample Types: Recombinant Protein
    Applications: Bioassay
  2. Secretomics reveals gelatinase substrates at the blood-brain barrier that are implicated in astroglial barrier function
    Authors: Burmeister, M;Fraunenstein, A;Kahms, M;Arends, L;Gerwien, H;Deshpande, T;Kuhlmann, T;Gross, CC;Naik, VN;Wiendl, H;Klingauf, J;Meissner, F;Sorokin, L;
    Science advances
    Species: Mouse
    Sample Types: Recombinant Protein
    Applications: Bioassay
  3. Doxycycline Attenuates Doxorubicin-Induced Cardiotoxicity by Improving Myocardial Energy Metabolism in Rats
    Authors: D Dantas, AG Pereira, ASS Fujimori, APD Ribeiro, CCV de Almeida, MG Monte, CR Corrêa, AA Fernandes, SGZ Bazan, PS Azevedo, MF Minicucci, SAR de Paiva, LAM Zornoff, BF Polegato
    Journal of cardiovascular development and disease, 2022-08-08;9(8):.
    Species: N/A
    Sample Types: Reference Standard
    Applications: Zymography
  4. Matrix metallopeptidase expression and modulation by transforming growth factor-beta1 in equine endometrosis
    Authors: A Szóstek-Mi, M S?owi?ska, J Pacewicz, DJ Skarzynski, K Okuda
    Sci Rep, 2020-01-24;10(1):1119.
    Species: Mouse
    Sample Types: Protein
    Applications: Reference Standard
  5. Matrix Metalloproteinase-3 is Key Effector of TNF-alpha-Induced Collagen Degradation in Skin
    Authors: U Mirastschi, B Lupše, K Maedler, B Sarma, A Radtke, G Belge, M Dorsch, D Wedekind, LJ McCawley, G Boehm, U Zier, K Yamamoto, S Kelm, MS Ågren
    Int J Mol Sci, 2019-10-22;20(20):.
    Species: Mouse
    Sample Types: Protein
    Applications: Zymography Standard
  6. MMP Inhibition Preserves Integrin Ligation and FAK Activation to Induce Survival and Regeneration in RGCs Following Optic Nerve Damage
    Authors: PM D'Onofrio, AP Shabanzade, BK Choi, M Bähr, PD Koeberle
    Invest. Ophthalmol. Vis. Sci., 2019-02-01;60(2):634-649.
    Species: Rat
    Sample Types: Recombinant Protein
    Applications: Western Blot
  7. Loss of Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1) in the Diabetic Retina: Role of Matrix Metalloproteinases
    Authors: RS Eshaq, NR Harris
    Invest. Ophthalmol. Vis. Sci., 2019-02-01;60(2):748-760.
    Species: Rat
    Sample Types: Whole Cells
    Applications: Bioassay
  8. Tissue inhibitor of metalloproteinases 1 enhances rod survival in the rd1 mouse retina
    Authors: HS Kim, A Vargas, YS Eom, J Li, KL Yamamoto, CM Craft, EJ Lee
    PLoS ONE, 2018-05-09;13(5):e0197322.
    Species: Mouse
    Sample Types: Recombinant Protein
    Applications: Zymography
  9. Matrix Metalloproteinase-Mediated Blood-Brain Barrier Dysfunction in Epilepsy
    Authors: RG Rempe, AMS Hartz, ELB Soldner, BS Sokola, SR Alluri, EL Abner, RJ Kryscio, A Pekcec, J Schlichtig, B Bauer
    J. Neurosci., 2018-04-09;0(0):.
    Species: Rat
    Sample Types:
    Applications: Zymography
  10. Inhibition of Matrix Metalloproteinase 9 Enhances Rod Survival in the S334ter-line3 Retinitis Pigmentosa Model
    PLoS ONE, 2016-11-28;11(11):e0167102.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Zymography
  11. Pamidronate Attenuates Oxidative Stress and Energetic Metabolism Changes but Worsens Functional Outcomes in Acute Doxorubicin-Induced Cardiotoxicity in Rats
    Authors: Bertha Furlan Polegato
    Cell. Physiol. Biochem., 2016-11-25;40(4):431-442.
    Applications: ELISA (Standard)
  12. Tumor-induced pressure in the bone microenvironment causes osteocytes to promote the growth of prostate cancer bone metastases.
    Authors: Sottnik J, Dai J, Zhang H, Campbell B, Keller E
    Cancer Res, 2015-04-08;75(11):2151-8.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  13. Cannabinoid CB2 receptors regulate central sensitization and pain responses associated with osteoarthritis of the knee joint.
    Authors: Burston, James J, Sagar, Devi Ran, Shao, Pin, Bai, Mingfeng, King, Emma, Brailsford, Louis, Turner, Jenna M, Hathway, Gareth J, Bennett, Andrew J, Walsh, David A, Kendall, David A, Lichtman, Aron, Chapman, Victoria
    PLoS ONE, 2013-11-25;8(11):e80440.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Zymography
  14. G protein-coupled receptor kinase 6 deficiency promotes angiogenesis, tumor progression, and metastasis.
    Authors: Raghuwanshi S, Smith N, Rivers E, Thomas A, Sutton N, Hu Y, Mukhopadhyay S, Chen X, Leung T, Richardson R
    J Immunol, 2013-04-15;190(10):5329-36.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: ELISA (Control)
  15. In vivo processing of CXCL5 (LIX) by matrix metalloproteinase (MMP)-2 and MMP-9 promotes early neutrophil recruitment in IL-1beta-induced peritonitis.
    Authors: Song J, Wu C, Zhang X, Sorokin L
    J Immunol, 2012-12-07;190(1):401-10.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Cell Culture
  16. Ocular neovascularization caused by herpes simplex virus type 1 infection results from breakdown of binding between vascular endothelial growth factor A and its soluble receptor.
    Authors: Suryawanshi A, Mulik S, Sharma S
    J. Immunol., 2011-02-16;186(6):3653-65.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Enzyme Assay
  17. The selective MMP-12 inhibitor, AS111793 reduces airway inflammation in mice exposed to cigarette smoke.
    Authors: Le Quement C, Guenon I, Gillon JY, Valenca S, Cayron-Elizondo V, Lagente V, Boichot E
    Br. J. Pharmacol., 2008-05-19;154(6):1206-15.
    Applications: Zymography
  18. A GMCSF and IL-15 fusokine leads to paradoxical immunosuppression in vivo via asymmetrical JAK/STAT signaling through the IL-15 receptor complex.
    Authors: Rafei M, Wu JH, Annabi B, Lejeune L, Francois M, Galipeau J
    Blood, 2006-11-02;109(5):2234-42.
    Species: Mouse
    Sample Types: N/A, Whole Cells
    Applications: Bioassay, Western Blot
  19. Activity-dependent release of precursor nerve growth factor, conversion to mature nerve growth factor, and its degradation by a protease cascade.
    Authors: Bruno MA, Cuello AC
    Proc. Natl. Acad. Sci. U.S.A., 2006-04-17;103(17):6735-40.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Bioassay

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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