Human Pro-MMP-13 Quantikine ELISA Kit

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PDM1300
SMP1300
DM1300
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MMP-13 ELISA Kit ELISA
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Citations (28)
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Human Pro-MMP-13 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)
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.

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.
Human Pro-MMP-13 ELISA Linearity

Data Examples

Human Pro-MMP-13 ELISA Standard Curve

Product Datasheets

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
⚠ 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. 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 for Human Pro-MMP-13 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.

28 Citations: Showing 1 - 10
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  1. 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 Types: Cell Culture Supernates
  2. 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 Types: Synovial Lavage Fluid
  3. Hypoxia-inducible factor-l? mediates aggrecan and collagen ? expression via NOTCH1 signaling in nucleus pulposus cells during intervertebral disc degeneration
    Authors: Z Liu, C Li, X Meng, Y Bai, J Qi, J Wang, Q Zhou, W Zhang, X Zhang
    Biochem. Biophys. Res. Commun., 2017;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. 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 Types: Cell Culture Supernates
  5. 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
  6. 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 Types: Cell Culture Supernates
  7. 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 Types: Cell Culture Supernates
  8. 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
  9. 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
  10. 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
  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 Types: Cell Culture Supernates
  12. 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 Types: Cell Culture Supernates
  13. 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 Types: Cell Culture Supernates
  14. 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
  15. 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 Types: Cell Culture Supernates
  16. 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 Types: 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 Types: Cell Culture Supernates
  18. 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 Types: Cell Culture Supernates
  19. 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
  20. 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 Types: Cell Culture Supernates
  21. 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 Types: Synovial Fluid
  22. 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 Types: Cell Culture Supernates
  23. 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 Types: Cell Culture Supernates
  24. 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 Types: Cell Culture Supernates
  25. 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 Types: Synovial Fluid
  26. 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 Types: Cell Culture Supernates
  27. 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 Types: Cell Culture Supernates
  28. 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

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