Recombinant Mouse/Rat MMP-2 Protein, CF Summary
Product Specifications
Ile34-Cys662
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.
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.
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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 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
- Dilute rm/rMMP-2 to 40 µg/mL with 1 mM APMA in Assay Buffer.
- Incubate at 37 °C for 2 hours to activate rm/rMMP-2.
- Dilute activated rm/rMMP-2 to 0.2 ng/µL in Assay Buffer.
- Dilute Substrate to 80 µM in Assay Buffer.
- 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.
- 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:- rm/rMMP-2: 0.010 µg
- Substrate: 40 µM
Reconstitution Calculator
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.
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.
20
Citations: Showing 1 - 10
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Preclinical development of SGN-CD47M: Protease-activated antibody technology enables selective tumor targeting of the innate immune checkpoint receptor CD47
Authors: Levengood, MR;Carosino, CM;Zhang, X;Lucas, S;Ortiz, DJ;Westendorf, L;Chin, AP;Martin, AD;Wong, A;Hengel, SM;Sun, H;Zeng, W;Yumul, R;Dominguez, MM;Chen, Y;Zheng, JH;Karlsson, CAB;Trang, VH;Senter, PD;Gardai, SJ;
Molecular cancer therapeutics
Species: N/A
Sample Types: Antibody
Applications: Bioassay -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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) -
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 -
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 -
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) -
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 -
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 -
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 -
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 -
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
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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.
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