Human MMP-2 DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human MMP-2. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
Background: MMP-2
The matrix metalloproteinases (MMPs) consist of 24 known human zinc proteases with essential roles in breaking down components of the extracellular matrix (ECM). Additional MMP substrates include cytokines, chemokines, growth factors and binding proteins, cell/cell adhesion molecules, and other proteinases. With a few exceptions, MMPs share common structural motifs including a pro-peptide domain, a catalytic domain, a hinge region, and a hemopexin-like domain. Synthesized as pro-enzymes, most MMPs are secreted before conversion to their active form. MMP activities are modulated on several levels including transcription, pro-enzyme activation, or by their endogenous inhibitors, tissue inhibitors of metalloproteinases (TIMPs). A subset of MMPs are associated with membranes and designated as membrane-type metalloproteinases (MT-MMP).
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human MMP-2 DuoSet ELISA
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.
21
Citations: Showing 1 - 10
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Anti-Inflammatory and Antioxidant Properties of a New Mixture of Vitamin C, Collagen Peptides, Resveratrol, and Astaxanthin in Tenocytes: Molecular Basis for Future Applications in Tendinopathies
Authors: Marzagalli, M;Battaglia, S;Raimondi, M;Fontana, F;Cozzi, M;Ranieri, FR;Sacchi, R;Curti, V;Limonta, P;
Mediators of inflammation
Species: Human
Sample Types: Cell Culture Supernates
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Integrative Analyses of Circulating Proteins and Metabolites Reveal Sex Differences in the Associations with Cardiac Function among DCM Patients
Authors: Hannemann, A;Ameling, S;Lehnert, K;Dörr, M;Felix, SB;Nauck, M;Al-Noubi, MN;Schmidt, F;Haas, J;Meder, B;Völker, U;Friedrich, N;Hammer, E;
International journal of molecular sciences
Species: Human
Sample Types: Plasma
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Diagnostic Utility of Selected Matrix Metalloproteinases (MMP-2, MMP-3, MMP-11, MMP-26), HE4, CA125 and ROMA Algorithm in Diagnosis of Ovarian Cancer
Authors: Kicman, A;Gacuta, E;Kulesza, M;B?dkowska, EG;Marecki, R;Klank-Soko?owska, E;Knapp, P;Niczyporuk, M;?awicki, S;
International journal of molecular sciences
Species: Human
Sample Types: Plasma
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IL-17A in Human Liver: Significant Source of Inflammation and Trigger of Liver Fibrosis Initiation.
Authors: Kartasheva-Ebertz D, Gaston J, Lair-Mehiri L, Mottez E, Buivan T, Massault P, Scatton O, Gaujoux S, Vaillant J, Pol S, Lagaye S
Int J Mol Sci, 2022-08-29;23(17):.
Species: Human
Sample Types: Tissue Culture Supernates
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Mg-based materials diminish tumor spreading and cancer metastases
Authors: P Globig, R Madurawala, R Willumeit-, F Martini, E Mazzoni, BJC Luthringer
Bioactive materials, 2022-05-10;19(0):594-610.
Species: Human
Sample Types: Cell Culture Supernates
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Role of Betaglycan in TGF-beta Signaling and Wound Healing in Human Endometriotic Epithelial Cells and in Endometriosis
Authors: AN Mwaura, MA Riaz, JB Maoga, E Mecha, COA Omwandho, G Scheiner-B, I Meinhold-H, L Konrad
Biology, 2022-03-26;11(4):.
Species: Human
Sample Types: Cell Culture Supernates
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A microphysiological model of human trophoblast invasion during implantation
Authors: JY Park, S Mani, G Clair, HM Olson, VL Paurus, CK Ansong, C Blundell, R Young, J Kanter, S Gordon, AY Yi, M Mainigi, DD Huh
Nature Communications, 2022-03-15;13(1):1252.
Species: Human
Sample Types: Cell Culture Supernates
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Antibody microarray analysis of amniotic fluid proteomes in women with cervical insufficiency and short cervix, and their association with pregnancy latency length
Authors: S Hong, KH Park, YE Lee, JE Lee, YM Kim, E Joo, I Cho
PLoS ONE, 2022-02-07;17(2):e0263586.
Species: Human
Sample Types: Amniotic Fluid
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MMPs and TIMPs levels are correlated with anthropometric parameters, blood pressure, and endothelial function in obesity
Authors: S Boumiza, K Chahed, Z Tabka, MP Jacob, X Norel, G Ozen
Scientific Reports, 2021-10-08;11(1):20052.
Species: Human
Sample Types: Plasma
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The Value of Fetuin-A as a Predictor to Identify Takotsubo Patients at Risk of Cardiovascular Events
Authors: A Topf, M Mirna, N Bacher, V Paar, C Edlinger, LJ Motloch, S Gharibeh, M Bannehr, UC Hoppe, M Lichtenaue
Journal of cardiovascular development and disease, 2021-10-05;8(10):.
Species: Human
Sample Types: Serum
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Microfluidic Single-Cell Proteomics Assay Chip: Lung Cancer Cell Line Case Study
Authors: Y Jung, M Son, YR Nam, J Choi, JR Heath, S Yang
Micromachines, 2021-09-23;12(10):.
Species: Human
Sample Types: Cell Lysates
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ADAMTS-5 Decreases in Coronary Arteries and Plasma from Patients with Coronary Artery Disease
Authors: Z Wang, D Ye, J Ye, M Wang, J Liu, H Jiang, Y Xu, J Zhang, J Chen, J Wan
Dis. Markers, 2019-12-16;2019(0):6129748.
Species: Human
Sample Types: Plasma
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Immune and Inflammatory Proteins in Cord Blood as Predictive Biomarkers of Retinopathy of Prematurity in Preterm Infants
Authors: YJ Park, SJ Woo, YM Kim, S Hong, YE Lee, KH Park
Invest. Ophthalmol. Vis. Sci., 2019-09-03;60(12):3813-3820.
Species: Human
Sample Types: Plasma
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Glycosaminoglycans influence enzyme activity of MMP2 and MMP2/TIMP3 complex formation - Insights at cellular and molecular level
Authors: G Ruiz-Gómez, S Vogel, S Möller, MT Pisabarro, U Hempel
Sci Rep, 2019-03-20;9(1):4905.
Species: Human
Sample Types: Cell Culture Supernates
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Fibrin Glue Enhances Adipose-Derived Stromal Cell Cytokine Secretion and Survival Conferring Accelerated Diabetic Wound Healing
Authors: U Hopfner, MM Aitzetmuel, P Ne beta bach, MS Hu, HG Machens, ZN Maan, D Duscher
Stem Cells Int, 2018-12-19;2018(0):1353085.
Species: Human
Sample Types: Cell Culture Supernates
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Novel oncogenic and chemoresistance-inducing functions of resistin in ovarian cancer cells require miRNAs-mediated induction of epithelial-to-mesenchymal transition
Authors: L Qiu, GF Zhang, L Yu, HY Wang, XJ Jia, TJ Wang
Sci Rep, 2018-08-21;8(1):12522.
Species: Human
Sample Types: Cell Culture Supernates
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Processed eggshell membrane powder regulates cellular functions and increase MMP-activity important in early wound healing processes
Authors: TT Vuong, SB Rønning, TAE Ahmed, K Brathagen, V Høst, MT Hincke, HP Suso, ME Pedersen
PLoS ONE, 2018-08-06;13(8):e0201975.
Species: Human
Sample Types: Cell Culture Supernates
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SH003 reverses drug resistance by blocking signal transducer and activator of transcription 3 (STAT3) signaling in breast cancer cells
Authors: HS Seo, JM Ku, HJ Lee, JK Woo, C Cheon, M Kim, BH Jang, YC Shin, SG Ko
Biosci. Rep., 2017-11-15;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Periodontitis and type 2 diabetes among women with previous gestational diabetes: epidemiological and immunological aspects in a follow-up of three years
Authors: RP Esteves Li, LO Cota, TA Silva, SC Cortelli, JR Cortelli, FO Costa
J Appl Oral Sci, 2017-03-01;25(2):130-139.
Species: Human
Sample Types: Saliva
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TLR ligands stimulation protects MSC from NK killing.
Authors: Giuliani M, Bennaceur-Griscelli A, Nanbakhsh A, Oudrhiri N, Chouaib S, Azzarone B, Durrbach A, Lataillade J
Stem Cells, 2014-01-01;32(1):290-300.
Species: Human
Sample Types: Cell Culture Supernates
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Stromal fibroblasts in the microenvironment of gastric carcinomas promote tumor metastasis via upregulating TAGLN expression.
Authors: Yu B, Chen X, Li J, Qu Y, Su L, Peng Y, Huang J, Yan J, Yu Y, Gu Q, Zhu Z, Liu B
BMC Cell Biol, 2013-03-19;14(0):17.
Species: Human
Sample Types: Cell Culture Supernates
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