Human Total MMP-3 DuoSet ELISA

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Ancillary Products Available
Human Total MMP-3 ELISA Standard Curve
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Product Details
Citations (19)
Supplemental Products
Reviews (2)

Human Total MMP-3 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* 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 TotalMMP-3. 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

Human Total MMP-3 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: MMP-3

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).

Long Name:
Matrix Metalloproteinase 3
Entrez Gene IDs:
4314 (Human); 17392 (Mouse)
Alternate Names:
CHDS6; EC 3.4.24; EC; matrix metallopeptidase 3 (stromelysin 1, progelatinase); matrix metalloproteinase 3 (stromelysin 1, progelatinase); Matrix metalloproteinase-3; MGC126102; MGC126103; MMP3; MMP-3; proteoglycanase; SL-1; STMY; STMY1MGC126104; STR1; Stromelysin 1; stromelysin-1; transin-1

Assay Procedure


Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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 Total MMP-3 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.

19 Citations: Showing 1 - 10
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  1. Development of an Electrochemical CCL5 Chemokine Immunoplatform for Rapid Diagnosis of Multiple Sclerosis
    Authors: S Guerrero, E Sánchez-Ti, L Agüí, A González-C, P Yáñez-Sede, JM Pingarrón
    Biosensors, 2022;12(8):.
    Species: Human
    Sample Types: Serum
  2. 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;11(4):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Nobiletin Decreases Inflammatory Mediator Expression in Tumor Necrosis Factor-Stimulated Human Periodontal Ligament Cells
    Authors: Y Hosokawa, I Hosokawa, K Ozaki
    Mediators of Inflammation, 2021;2021(0):5535844.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Differential inflammation-mediated function of prokineticin 2 in the synovial fibroblasts of patients with rheumatoid arthritis compared with osteoarthritis
    Authors: K Noda, B Dufner, H Ito, K Yoshida, G Balboni, RH Straub
    Scientific Reports, 2021;11(1):18399.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Targeting of Janus Kinases Limits Pro-Inflammatory but Also Immunosuppressive Circuits in the Crosstalk between Synovial Fibroblasts and Lymphocytes
    Authors: N Yao, T Tretter, P Kvacskay, W Merkt, N Blank, HM Lorenz, LO Tykocinski
    Biomedicines, 2021;9(10):.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. 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;11(1):20052.
    Species: Human
    Sample Types: Plasma
  7. Increase of aerobic glycolysis mediated by activated T helper cells drives synovial fibroblasts towards an inflammatory phenotype: new targets for therapy?
    Authors: P Kvacskay, N Yao, JH Schnotz, R Scarpone, RA Carvalho, KD Klika, W Merkt, T Tretter, HM Lorenz, LO Tykocinski
    Arthritis Research & Therapy, 2021;23(1):56.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. The Polymethoxy Flavonoid Sudachitin Inhibits Interleukin-1&beta-Induced Inflammatory Mediator Production in Human Periodontal Ligament Cells
    Authors: Y Hosokawa, I Hosokawa, K Ozaki, T Matsuo
    BioMed Research International, 2021;2021(0):8826586.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Evaluation of the post-treatment anti-inflammatory capacity of osteoarthritic chondrocytes: An in�vitro study using baicalein
    Authors: CC Wu, YR Chen, DH Lu, LH Hsu, KC Yang, S Sumi
    Regen Ther, 2020;14(0):177-183.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. 1,25(OH)2D3 and dexamethasone additively suppress synovial fibroblast activation by CCR6+ T helper memory cells and enhance the effect of tumor necrosis factor alpha blockade
    Authors: W Dankers, C González-L, N Davelaar, PS Asmawidjaj, AMC Mus, JMW Hazes, EM Colin, E Lubberts
    Arthritis Res. Ther., 2018;20(1):212.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Serum metabolomic profiling predicts synovial gene expression in rheumatoid arthritis
    Authors: R Narasimhan, R Coras, SB Rosenthal, SR Sweeney, A Lodi, S Tiziani, D Boyle, A Kavanaugh, M Guma
    Arthritis Res. Ther., 2018;20(1):164.
    Species: Human
    Sample Types: Serum
  12. Functionally distinct disease-associated fibroblast subsets in rheumatoid arthritis
    Authors: F Mizoguchi, K Slowikowsk, K Wei, JL Marshall, DA Rao, SK Chang, HN Nguyen, EH Noss, JD Turner, BE Earp, PE Blazar, J Wright, BP Simmons, LT Donlin, GD Kalliolias, SM Goodman, VP Bykerk, LB Ivashkiv, JA Lederer, N Hacohen, PA Nigrovic, A Filer, CD Buckley, S Raychaudhu, MB Brenner
    Nat Commun, 2018;9(1):789.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Role of MMP-1 (-519A/G, -1607 1G/2G), MMP-3 (Lys45Glu), MMP-7 (-181A/G), and MMP-12 (-82A/G) Variants and Plasma MMP Levels on Obesity-Related Phenotypes and Microvascular Reactivity in a Tunisian Population
    Authors: S Boumiza, S Bchir, H Ben Nasr, A Abbassi, MP Jacob, X Norel, Z Tabka, K Chahed
    Dis. Markers, 2017;2017(0):6198526.
    Species: Human
    Sample Types: Plasma
  14. Human cytomegalovirus interleukin-10 enhances matrigel invasion of MDA-MB-231 breast cancer cells
    Authors: CA Valle Oseg, JV Spencer
    Cancer Cell Int, 2017;17(0):24.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Dimethyl fumarate protection against collagen II degradation.
    Authors: Li Y, Tang J, Hu Y
    Biochem Biophys Res Commun, 2014;454(2):257-61.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. 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;32(1):290-300.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Matrix metalloproteinase-3 and intracranial arterial dolichoectasia.
    Authors: Pico F, Jacob MP, Labreuche J, Soufir N, Touboul PJ, Benessiano J, Cambien F, Grandchamp B, Michel JB, Amarenco P
    Ann. Neurol., 2010;67(4):508-15.
    Species: Human
    Sample Types: Plasma
  18. Serum levels of sex steroid hormones and matrix metalloproteinases after intra-articular glucocorticoid treatment in female patients with rheumatoid arthritis.
    Authors: Weitoft T, Larsson A, Ronnblom L
    Ann. Rheum. Dis., 2008;67(3):422-4.
    Species: Human
    Sample Types: Serum
  19. Altered expression of MicroRNA in synovial fibroblasts and synovial tissue in rheumatoid arthritis.
    Authors: Stanczyk J, Pedrioli DM, Brentano F, Sanchez-Pernaute O, Kolling C, Gay RE, Detmar M, Gay S, Kyburz D
    Arthritis Rheum., 2008;58(4):1001-9.
    Species: Human
    Sample Types: Cell Culture Supernates


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Reviews for Human Total MMP-3 DuoSet ELISA

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Human Total MMP-3 DuoSet ELISA
By Nicole Stone on 01/17/2019
Sample Tested: Human cell conditioned medium

Great product, very easy to use! Produced a great standard curve.

Human Total MMP-3 DuoSet ELISA
By Sophie Millar on 01/11/2019
Sample Tested: HASMC human aortic smooth muscle cells,Human aortic endothelial cells