Human Total MMP-3 Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
PDMP300
SMP300
DMP300
Control Products Available
Human Total MMP-3 ELISA Standard Curve
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Product Details
Procedure
Citations (39)
FAQs
Supplemental Products
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Human Total MMP-3 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (10 uL), Heparin Plasma (10 uL)
Sensitivity
0.045 ng/mL
Assay Range
0.2 - 10 ng/mL (Cell Culture Supernates, Serum, Heparin Plasma)
Specificity
Natural and recombinant human total MMP-3
Cross-reactivity
Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity not tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human Total MMP-3 Immunoassay is a 4.5 hour solid phase immunoassay designed to measure total MMP-3 (pro- and active MMP-3) in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human Pro-MMP-3 and antibodies raised against the recombinant factor. Both antibodies also recognize recombinant human active MMP-3. Natural human MMP-3 showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards, indicating that this kit can be used to determine relative levels of natural human MMP-3.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 0.962 3.21 5.87 0.994 3.08 5.7
Standard Deviation 0.062 0.197 0.332 0.085 0.217 0.449
CV% 6.4 6.1 5.7 8.6 7 7.9

Recovery

The recovery of MMP-3 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 96 87-104
Heparin Plasma (n=5) 92 85-106
Serum (n=5) 90 86-96

Linearity

To assess the linearity of the assay, samples containing high concentrations of MMP-3 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Total MMP-3 ELISA Linearity

Data Examples

Human Total MMP-3 ELISA Standard Curve

Product Datasheets

Preparation and Storage

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

Background: MMP-3

Matrix metalloproteinases (MMPs), also called matrixins, constitute a family of zinc and calcium dependent endopeptidases that function in the breakdown of extracellular matrix (ECM). They play an important role in many normal physiological processes such as embryonic development, morphogenesis, reproduction and tissue remodeling (1). They also participate in many pathological processes such as arthritis, cancer and cardiovascular disease (2). While the amounts of newly synthesized MMPs are regulated mainly at the levels of transcription, the proteolytic activities of existing MMPs are controlled through both the activation of proenzymes or zymogens and the inhibition of active enzymes by endogenous inhibitors, alpha -macroglobulins and tissue inhibitors of metalloproteinases (TIMPs). 

MMP-3 (also referred to as stromelysin-1) may be expressed in fibroblasts, chondrocytes, endothelial cells, macrophages, vascular smooth muscle cells, osteoblasts, and keratinocytes in response to appropriate stimuli (3). Various agents regulate its biosynthesis. Inflammatory cytokines such as IL-1 and TNF-alpha, epidermal growth factor, platelet-derived growth factor, phorbol and oncogenic cellular transformation are the inductive agents. In comparison, retinoic acid, glucocorticoids, estrogen, progesterone and TGF-beta suppress MMP-3 synthesis. 
MMP-3 is secreted from the cells as a proenzyme. The proenzyme has been shown to stimulate plasminogen activation (4). The N-terminal pro-domain contains the cysteine switch motif conserved in MMPs that maintains MMP-3 in the latent state (5). Activation of the proenzyme results in the removal of the pro-domain. MMP-3 activation can be achieved in vitro by proteases such as itself, chyrotrypsin, neutrophil elastase and plasma kallikrein, and by mercury compounds (3). The resulting active enzyme consists of a catalytic domain with a zinc-binding motif conserved in metzincins (6,7). A short hinge peptide links the catalytic domain to the C-terminal hemopexin-like domain. The active MMP-3 is capable of cleaving types III, IV, IX and X collagen, aggrecan, fibronectin, laminin, IGFBP-3, serpins, and IL-1 beta. The active enzyme also activates proMMP-1, -8, -9, and -13. Therefore, it is suggested that MMP-3 may participate in physiological matrix turnover and pathological destruction of the tissue. For example, MMP-3 is required for the generation of a macrophage chemoattractant in a model of herniated disc resorption (8).

Long Name:
Matrix Metalloproteinase 3
Entrez Gene IDs:
4314 (Human); 17392 (Mouse)
Alternate Names:
CHDS6; EC 3.4.24; EC 3.4.24.17; 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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 100 µL Standard, Control, or Sample
  6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Total MMP-3 Quantikine ELISA Kit

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.

39 Citations: Showing 1 - 10
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  1. Activity of fibroblast-like synoviocytes in rheumatoid arthritis was impaired by dickkopf-1 targeting siRNA
    Authors: YY Liu, SY Wang, YN Li, WJ Bian, LQ Zhang, YH Li, L Long, X Liu, XW Zhang, ZG Li
    Chin. Med. J., 2020;0(6):679-686.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Basilar Artery Tortuosity Is Associated With White Matter Hyperintensities by TIMP-1
    Authors: DP Zhang, YF Peng, HL Zhang, JG Ma, M Zhao, S Yin, TT Wei
    Front Neurosci, 2019;13(0):836.
    Species: Human
    Sample Types: Plasma
  3. PhosphoLipid transfer protein (PLTP) exerts a direct pro-inflammatory effect on rheumatoid arthritis (RA) fibroblasts-like-synoviocytes (FLS) independently of its lipid transfer activity
    Authors: R Audo, V Deckert, CI Daien, H Che, J Elhmioui, S Lemaire, JP Pais de Ba, C Desrumaux, B Combe, M Hahne, L Lagrost, J Morel
    PLoS ONE, 2018;13(3):e0193815.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Nesfatin-1 and visfatin expression is associated with reduced atherosclerotic disease risk in patients with rheumatoid arthritis
    Authors: C Robinson, L Tsang, A Solomon, AJ Woodiwiss, S Gunter, M Mer, HC Hsu, M Gomes, GR Norton, AME Millen, PH Dessein
    Peptides, 2018;102(0):31-37.
    Species: Human
    Sample Types: Plasma
  5. Increased levels of circulating MMP3 correlate with severe rejection in face transplantation
    Authors: B Kollar, A Shubin, TJ Borges, S Tasigiorgo, TS Win, CG Lian, ST Dillon, X Gu, I Wyrobnik, GF Murphy, B Pomahac, TA Libermann, LV Riella
    Sci Rep, 2018;8(1):14915.
    Species: Human
    Sample Types: Serum
  6. Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation
    Authors: CH Hulme, EL Wilson, HR Fuller, S Roberts, JB Richardson, P Gallacher, MJ Peffers, SL Shirran, CH Botting, KT Wright
    Arthritis Res. Ther., 2018;20(1):87.
    Species: Human
    Sample Types: Synovial Fluid
  7. Late mitral restenosis after percutaneous commissurotomy: Predictive value of inflammation and extracellular matrix remodeling biomarkers
    Authors: R Mechmeche, A Zaroui, S Aloui, M Boukhris, M Allal-Elas, N Kaabachi, B Zouari
    Heart Lung, 2017;0(0):.
    Species: Human
    Sample Types: Serum
  8. CX3CL1 promotes MMP-3 production via the CX3CR1, c-Raf, MEK, ERK, and NF-?B signaling pathway in osteoarthritis synovial fibroblasts
    Authors: SM Hou, CH Hou, JF Liu
    Arthritis Res. Ther., 2017;19(1):282.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Serum bone-turnover biomarkers are associated with the occurrence of peripheral and axial arthritis in psoriatic disease: a prospective cross-sectional comparative study
    Authors: DR Jadon, R Sengupta, A Nightingal, H Lu, J Dunphy, A Green, JT Elder, RP Nair, E Korendowyc, MA Lindsay, NJ McHugh
    Arthritis Res. Ther., 2017;19(1):210.
    Species: Human
    Sample Types: Serum
  10. Exosomes in Human Breast Milk Promote EMT
    Authors: W Qin, Y Tsukasaki, S Dasgupta, N Mukhopadhy, M Ikebe, ER Sauter
    Clin Cancer Res, 2016;0(0):.
    Species: Human
    Sample Types: Cell Lysates
  11. Concomitant elevations of MMP-9, NGAL, proMMP-9/NGAL and neutrophil elastase in serum of smokers with chronic obstructive pulmonary disease
    J. Cell. Mol. Med, 2016;0(0):.
    Species: Human
    Sample Types: Serum
  12. Galectin-3 Induces a Pro-degradative/inflammatory Gene Signature in Human Chondrocytes, Teaming Up with Galectin-1 in Osteoarthritis Pathogenesis
    Sci Rep, 2016;6(0):39112.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Metallothionein-3 Increases Triple-Negative Breast Cancer Cell Invasiveness via Induction of Metalloproteinase Expression.
    Authors: Kmiecik A, Pula B, Suchanski J, Olbromski M, Gomulkiewicz A, Owczarek T, Kruczak A, Ambicka A, Rys J, Ugorski M, Podhorska-Okolow M, Dziegiel P
    PLoS ONE, 2015;10(5):e0124865.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Exogenous IFN-beta regulates the RANKL-c-Fos-IFN-beta signaling pathway in the collagen antibody-induced arthritis model.
    Authors: Zhao R, Chen N, Zhou X, Miao P, Hu C, Qian L, Yu Q, Zhang J, Nie H, Chen X, Li P, Xu R, Xiao L, Zhang X, Liu J, Zhang D
    J Transl Med, 2014;12(1):330.
    Species: Human
    Sample Types: Serum
  15. Validation and quality control of ELISAs for the use with human saliva samples.
    Authors: Jaedicke KM, Taylor JJ, Preshaw PM
    J. Immunol. Methods, 2012;377(1):62-5.
    Species: Human
    Sample Types: Saliva
  16. Interleukin-6 and matrix metalloproteinase expression in the subretinal fluid during proliferative vitreoretinopathy: correlation with extent, duration of RRD and PVR grade.
    Authors: Symeonidis C, Papakonstantinou E, Androudi S, Tsaousis KT, Tsinopoulos I, Brazitikos P, Karakiulakis G, Diza E, Dimitrakos SA
    Cytokine, 2012;59(1):184-90.
    Species: Human
    Sample Types: Subretinal Fluid
  17. Interleukin-6 and the matrix metalloproteinase response in the vitreous during proliferative vitreoretinopathy.
    Authors: Symeonidis C, Papakonstantinou E, Androudi S, Rotsos T, Diza E, Brazitikos P, Karakiulakis G, Dimitrakos SA
    Cytokine, 2011;54(2):212-7.
    Species: Human
    Sample Types: Vitreous Humor
  18. Cyclooxygenase-2 inhibitor blocks the production of West Nile virus-induced neuroinflammatory markers in astrocytes.
    Authors: Verma S, Kumar M, Nerurkar VR
    J. Gen. Virol., 2011;92(0):507-15.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Proteolytic release of the receptor for advanced glycation end products from in vitro and in situ alveolar epithelial cells.
    Authors: Yamakawa N, Uchida T, Matthay MA, Makita K
    Am. J. Physiol. Lung Cell Mol. Physiol., 2011;300(4):L516-25.
    Species: Human
    Sample Types: Edema Fluid
  20. Primary human acute myelogenous leukemia cells release matrix metalloproteases and their inhibitors: release profile and pharmacological modulation.
    Authors: Reikvam H, Hatfield KJ, Oyan AM, Kalland KH, Kittang AO, Bruserud O
    Eur. J. Haematol., 2010;84(3):239-51.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Matrix metalloproteinase-9 and plasminogen activator inhibitor-1 are associated with right ventricular structure and function: the MESA-RV Study.
    Authors: Kawut SM, Barr RG, Johnson WC
    Biomarkers, 2010;15(8):731-8.
    Species: Human
    Sample Types: Plasma
  22. Circulating matrix metalloproteinase-3 and metalloproteinase-9 and tissue Doppler measures of diastolic dysfunction to risk stratify patients with systolic heart failure.
    Authors: Buralli S, Dini FL, Ballo P, Conti U, Fontanive P, Duranti E, Metelli MR, Marzilli M, Taddei S
    Am. J. Cardiol., 2010;105(6):853-6.
    Species: Human
    Sample Types: Plasma
  23. Soluble biomarkers of cartilage and bone metabolism in early proof of concept trials in psoriatic arthritis: effects of adalimumab versus placebo.
    Authors: van Kuijk AW, DeGroot J, Koeman RC
    PLoS ONE, 2010;5(9):e12556.
    Species: Human
    Sample Types: Serum
  24. Elevated serum matrix metalloproteinase-3 and -7 in H. pylori-related gastric cancer can be biomarkers correlating with a poor survival.
    Authors: Yeh YC, Sheu BS, Cheng HC, Wang YL, Yang HB, Wu JJ
    Dig. Dis. Sci., 2010;55(6):1649-57.
    Species: Human
    Sample Types: Serum
  25. High molecular weight hyaluronic acid inhibits IL-6-induced MMP production from human chondrocytes by up-regulating the ERK inhibitor, MKP-1.
    Authors: Hashizume M, Mihara M
    Biochem. Biophys. Res. Commun., 2010;403(2):184-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  26. Establishment of a matrix-associated transepithelial resistance invasion assay to precisely measure the invasive potential of synovial fibroblasts.
    Authors: Wunrau C, Schnaeker EM, Freyth K, Pundt N, Wendholt D, Neugebauer K, Hansen U, Pap T, Dankbar B
    Arthritis Rheum., 2009;60(9):2606-11.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. Inhibition of IL-1beta-mediated inflammatory responses by the IkappaB alpha super-repressor in human fibroblast-like synoviocytes.
    Authors: Lee YR, Kweon SH, Kwon KB, Park JW, Yoon TR, Park BH
    Biochem. Biophys. Res. Commun., 2009;378(1):90-4.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. PI3Kgamma regulates cartilage damage in chronic inflammatory arthritis.
    Authors: Hayer S, Pundt N, Peters MA, Wunrau C, Kuhnel I, Neugebauer K, Strietholt S, Zwerina J, Korb A, Penninger J, Joosten LA, Gay S, Ruckle T, Schett G, Pap T
    FASEB J., 2009;23(12):4288-98.
    Species: Human
    Sample Types: Cell Culture Supernates
  29. Serum levels of matrix metalloproteinases -1,-2,-3 and -9 in thoracic aortic diseases and acute myocardial ischemia.
    Authors: Karapanagiotidis GT, Antonitsis P, Charokopos N, Foroulis CN, Anastasiadis K, Rouska E, Argiriadou H, Rammos K, Papakonstantinou C
    J Cardiothorac Surg, 2009;4(0):59.
    Species: Human
    Sample Types: Serum
  30. Gender specific associations between matrix metalloproteinases and inflammatory markers in post myocardial infarction patients.
    Authors: Samnegard A, Hulthe J, Silveira A, Ericsson CG, Hamsten A, Eriksson P
    Atherosclerosis, 2009;202(2):550-6.
    Species: Human
    Sample Types: Serum
  31. Serum levels of MMP-3 and cathepsin K in patients with ankylosing spondylitis: effect of TNFalpha antagonist therapy.
    Authors: Wendling D, Cedoz JP, Racadot E
    Joint Bone Spine, 2008;75(5):559-62.
    Species: Human
    Sample Types: Serum
  32. Targeting a metalloprotease-PAR1 signaling system with cell-penetrating pepducins inhibits angiogenesis, ascites, and progression of ovarian cancer.
    Authors: Agarwal A, Covic L, Sevigny LM, Kaneider NC, Lazarides K, Azabdaftari G, Sharifi S, Kuliopulos A
    Mol. Cancer Ther., 2008;7(9):2746-57.
    Species: Human
    Sample Types: Ovarian Ascites Fluid
  33. Actinobacillus actinomycetemcomitans lipopolysaccharide regulates matrix metalloproteinase, tissue inhibitors of matrix metalloproteinase, and plasminogen activator production by human gingival fibroblasts: a potential role in connective tissue destruction.
    Authors: Bodet C, Andrian E, Tanabe S, Grenier D
    J. Cell. Physiol., 2007;212(1):189-94.
    Species: Human
    Sample Types: Cell Culture Supernates
  34. Borrelia burgdorferi-induced monocyte chemoattractant protein-1 production in vivo and in vitro.
    Authors: Zhao Z, McCloud B, Fleming R, Klempner MS
    Biochem. Biophys. Res. Commun., 2007;358(2):528-33.
    Species: Human
    Sample Types: Tissue Homogenates
  35. Advanced glycation end products increases matrix metalloproteinase-1, -3, and -13, and TNF-alpha in human osteoarthritic chondrocytes.
    Authors: Nah SS, Choi IY, Yoo B, Kim YG, Moon HB, Lee CK
    FEBS Lett., 2007;581(9):1928-32.
    Species: Human
    Sample Types: Cell Culture Supernates
  36. Advanced glycation end products decrease mesangial cell MMP-7: a role in matrix accumulation in diabetic nephropathy?
    Authors: McLennan SV, Kelly DJ, Schache M, Waltham M, Dy V, Langham RG, Yue DK, Gilbert RE
    Kidney Int., 2007;72(4):481-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  37. Gingival fibroblasts inhibit MMP-1 and MMP-3 activities in an ex-vivo artery model.
    Authors: Naveau A, Reinald N, Fournier B
    Connect. Tissue Res., 2007;48(6):300-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  38. Stimulation of matrix metalloproteinases by hyperosmolarity via a JNK pathway in human corneal epithelial cells.
    Authors: Li DQ, Chen Z, Song XJ, Luo L, Pflugfelder SC
    Invest. Ophthalmol. Vis. Sci., 2004;45(12):4302-11.
    Species: Human
    Sample Types: Cell Culture Supernates
  39. Antidiabetic PPAR gamma-activator rosiglitazone reduces MMP-9 serum levels in type 2 diabetic patients with coronary artery disease.
    Authors: Marx N, Froehlich J, Siam L, Ittner J, Wierse G, Schmidt A, Scharnagl H, Hombach V, Koenig W
    Arterioscler. Thromb. Vasc. Biol., 2003;23(2):283-8.
    Species: Human
    Sample Types: Serum

FAQs

  1. Can I use EDTA or citrate plasma as samples in Human Total MMP-3 Quantikine ELISA Kit (Catalog # DMP300)?

    • MMPs are a family of zinc and calcium dependent endopeptidases, and MMP­3 catalytic domain contains a zinc binding site. Chelating agents such as EDTA and Citrate can sequester metal ions from the catalytic domain causing distruption of the MMP3 structure. MMP3 may be denatured as a result and may compromise measurements or detecteability in the assay.

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Human Total MMP-3 Quantikine ELISA Kit
By Anonymous on 10/14/2016
Application: Sample Tested: Cell Culture Supernates