Human Pro-MMP-13 Quantikine ELISA Kit

  (27 citations)     
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Assay Procedure
Citations (27)
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  • 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)
  • Sensitivity
    21.3 pg/mL
  • Assay Range
    78.0 - 5,000 pg/mL (Cell Culture Supernates)
  • Specificity
    Natural and recombinant human pro-MMP-13
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC132 , Quantikine Immunoassay Control Set 837 for Human Pro-MMP-13 - Please Inquire
Product Summary
The Quantikine Human Pro-MMP-13 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure pro-MMP-13 in cell culture supernates. It contains NS0-expressed recombinant human pro-MMP-13 and antibodies raised against the recombinant factor. The Quantikine kit will not detect recombinant human active MMP-13 existing in either free or TIMP-bound form. Natural human pro-MMP-13 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 pro-MMP-13.

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
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 485 1462 3521 533 1561 3823
Standard Deviation 18.2 64 93 27.9 56.1 143
CV% 3.8 4.4 2.6 5.2 3.6 3.7

Recovery

The recovery of pro-MMP-13 spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=2) 101 86-112
Cell Culture Supernates (n=3) 98 85-111
Linearity
To assess the linearity of the assay, samples containing or spiked with high concentrations of pro-MMP-13 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 MMP-13 [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: MMP-13

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 13
  • Entrez Gene IDs:
    4322 (Human)
  • Alternate Names:
    CLG 3; CLG3EC 3.4.24; collagenase 3; EC 3.4.24.-; EC 3.4.24.22; EC 3.4.24.24; EC 3.4.24.35; EC 3.4.24.65; EC 3.4.24.7; MANDP1; matrix metallopeptidase 13 (collagenase 3); matrix metalloproteinase 13 (collagenase 3); Matrix metalloproteinase-13; MMP13; MMP-13
Related Research Areas
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. 50 µL Standard, Control, or Sample
  6.   Add 50 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:

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.

27 Citations: Showing 1 - 10
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Species
Sample Type
  1. Synovial inflammation plays a greater role in post-traumatic osteoarthritis compared to idiopathic osteoarthritis in the Hartley guinea pig knee
    Authors: NP Thomas, WJ Wu, BC Fleming, F Wei, Q Chen, L Wei
    BMC Musculoskelet Disord, 2017;18(1):556.
    Species: Guinea Pig
    Sample Type: Synovial Lavage Fluid
  2. Kr�ppel-Like Factor 2 Regulates Degradation of Type II Collagen by Suppressing the Expression of Matrix Metalloproteinase (MMP)-13
    Authors: Y Yuan, H Tan, P Dai
    Cell. Physiol. Biochem., 2017;42(6):2159-2168.
    Species: Human
    Sample Type: Cell Culture Supernates
  3. Atmospheric particulate matter2.5 promotes the migration and invasion of hepatocellular carcinoma cells
    Authors: Q Zhang, Q Luo, X Yuan, L Chai, D Li, J Liu, Z Lv
    Oncol Lett, 2017;13(5):3445-3450.
    Species: Human
    Sample Type: Cell Culture Supernates
  4. 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 Type: Cell Culture Supernates
  5. Anti-arthritic effects of magnolol in human interleukin 1β-stimulated fibroblast-like synoviocytes and in a rat arthritis model.
    Authors: Wang JH, Shih KS, Liou JP, Wu YW, Chang AS, Wang KL, Tsai CL, Yang CR
    PLoS ONE, 2012;7(2):e31368.
    Species: Human
    Sample Type: Cell Culture Supernates
  6. Central role of SREBP-2 in the pathogenesis of osteoarthritis.
    Authors: Kostopoulou F, Gkretsi V, Malizos KN, Iliopoulos D, Oikonomou P, Poultsides L, Tsezou A
    PLoS ONE, 2012;7(5):e35753.
    Species: Human
    Sample Type: Cell Culture Supernates
  7. 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 Type: Edema Fluid
  8. 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 Type: Cell Culture Supernates
  9. 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 Type: Cell Culture Supernates
  10. Interleukin-7 stimulates secretion of S100A4 by activating the JAK/STAT signaling pathway in human articular chondrocytes.
    Authors: Yammani RR, Long D, Loeser RF
    Arthritis Rheum., 2009;60(3):792-800.
    Species: Human
    Sample Type: Cell Culture Supernates
  11. 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 Type: Cell Culture Supernates
  12. 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 Type: Ovarian Ascites Fluid
  13. Inhibition of interleukin-1beta-induced matrix metalloproteinases 1 and 13 production in human osteoarthritic chondrocytes by prostaglandin D2.
    Authors: Zayed N, Afif H, Chabane N, Mfuna-Endam L, Benderdour M, Martel-Pelletier J, Pelletier JP, Motiani RK, Trebak M, Duval N, Fahmi H
    Arthritis Rheum., 2008;58(11):3530-40.
    Species: Human
    Sample Type: Cell Culture Supernates
  14. Prostaglandin E2 exerts catabolic effects in osteoarthritis cartilage: evidence for signaling via the EP4 receptor.
    Authors: Attur M, Al-Mussawir HE, Patel J, Kitay A, Dave M, Palmer G, Pillinger MH, Abramson SB
    J. Immunol., 2008;181(7):5082-8.
    Species: Human
    Sample Type: Cell Culture Supernates
  15. Amidic alginate hydrogel for nucleus pulposus replacement.
    Authors: Leone G, Torricelli P, Chiumiento A, Facchini A, Barbucci R
    J Biomed Mater Res A, 2008;84(2):391-401.
    Species: Human
    Sample Type: Cell Culture Supernates
  16. 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 Type: Cell Culture Supernates
  17. ROCK-II mediates colon cancer invasion via regulation of MMP-2 and MMP-13 at the site of invadopodia as revealed by multiphoton imaging.
    Authors: Vishnubhotla R, Sun S, Huq J, Bulic M, Ramesh A, Guzman G, Cho M, Glover SC
    Lab. Invest., 2007;87(11):1149-58.
    Species: Human
    Sample Type: Cell Culture Supernates
  18. Basic fibroblast growth factor stimulates matrix metalloproteinase-13 via the molecular cross-talk between the mitogen-activated protein kinases and protein kinase Cdelta pathways in human adult articular chondrocytes.
    Authors: Im HJ, Muddasani P, Natarajan V, Schmid TM, Block JA, Davis F, van Wijnen AJ, Loeser RF
    J. Biol. Chem., 2007;282(15):11110-21.
    Species: Human
    Sample Type: Synovial Fluid
  19. Alendronate-hydroxyapatite nanocomposites and their interaction with osteoclasts and osteoblast-like cells.
    Authors: Boanini E, Torricelli P, Gazzano M, Giardino R, Bigi A
    Biomaterials, 2007;29(7):790-6.
    Species: Human
    Sample Type: Cell Culture Supernates
  20. Calcineurin modulates the catabolic and anabolic activity of chondrocytes and participates in the progression of experimental osteoarthritis.
    Authors: Yoo SA, Park BH, Yoon HJ, Lee JY, Song JH, Kim HA, Cho CS, Kim WU
    Arthritis Rheum., 2007;56(7):2299-311.
    Species: Human
    Sample Type: Cell Culture Supernates
  21. Oncostatin M induces angiogenesis and cartilage degradation in rheumatoid arthritis synovial tissue and human cartilage cocultures.
    Authors: Fearon U, Mullan R, Markham T, Connolly M, Sullivan S, Poole AR, FitzGerald O, Bresnihan B, Veale DJ
    Arthritis Rheum., 2006;54(10):3152-62.
    Species: Human
    Sample Type: Cell Culture Supernates
  22. The potential of adiponectin in driving arthritis.
    Authors: Ehling A, Schaffler A, Herfarth H, Tarner IH, Anders S, Distler O, Paul G, Distler J, Gay S, Scholmerich J, Neumann E, Muller-Ladner U
    J. Immunol., 2006;176(7):4468-78.
    Species: Human
    Sample Type: Cell Culture Supernates
  23. Increase in production of matrix metalloproteinase 13 by human articular chondrocytes due to stimulation with S100A4: Role of the receptor for advanced glycation end products.
    Authors: Yammani RR, Carlson CS, Bresnick AR, Loeser RF
    Arthritis Rheum., 2006;54(9):2901-11.
    Species: Human
    Sample Type: Cell Culture Supernates
  24. Induction of host matrix metalloproteinases by Borrelia burgdorferi differs in human and murine lyme arthritis.
    Authors: Behera AK, Hildebrand E, Scagliotti J, Steere AC, Hu LT
    Infect. Immun., 2005;73(1):126-34.
    Species: Human
    Sample Type: Synovial Fluid
  25. The role of human HtrA1 in arthritic disease.
    Authors: Grau S, Richards PJ, Kerr B, Hughes C, Caterson B, Williams AS, Junker U, Jones SA, Clausen T, Ehrmann M
    J. Biol. Chem., 2005;281(10):6124-9.
    Species: Human
    Sample Type: Cell Culture Supernates
  26. Differential modulation of human melanoma cell metalloproteinase expression by alpha2beta1 integrin and CD44 triple-helical ligands derived from type IV collagen.
    Authors: Baronas-Lowell D, Lauer-Fields JL, Borgia JA, Sferrazza GF, Al-Ghoul M, Minond D, Fields GB
    J. Biol. Chem., 2004;279(42):43503-13.
    Species: Human
    Sample Type: Cell Culture Supernates
  27. 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 Type: Cell Culture Supernates
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Quantikine Immunoassay Control Set 837 for Human Pro-MMP-13

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