Mouse Total MMP-9 Quantikine ELISA Kit

(27 citations)
(1 Review)
  
  • 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
    0.014 ng/mL
  • Assay Range
    0.1 - 5 ng/mL (Cell Culture Supernates, Serum, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse MMP-9 (pro-, active, and TIMP-complexed)
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Mouse MMP-9 (total) Immunoassay is a 4.5 hour solid-phase ELISA designed to measure total mouse MMP-9 (pro-, active, and TIMP-complexed MMP-9) in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse MMP-9 and antibodies raised against the recombinant factor. Natural mouse MMP-9 showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative levels of natural mouse MMP-9.

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
Sample123123
n202020404040
Mean0.2390.7311.710.2110.61.61
Standard Deviation0.0240.0370.0970.020.0490.125
CV%8.45.15.79.68.17.8

Recovery

The recovery of mouse MMP-9 spiked to 3 levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 94 86-108
Linearity
To assess the linearity of the assay, samples containing high concentrations of mouse MMP-9 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse Total MMP-9 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: MMP-9

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 9
    • Entrez Gene IDs
      4318 (Human); 17395 (Mouse); 81687 (Rat);
    • Alternate Names
      92 kDa gelatinase; 92 kDa type IV collagenase; CLG4B; EC 3.4.24; EC 3.4.24.35; Gelatinase B; GELB; macrophage gelatinase; MANDP2; matrix metallopeptidase 9; matrix metalloproteinase 9; matrix metalloproteinase-9; MMP9; MMP-9; type V collagenase;
    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. 50 µL Assay Diluent
    4.   Add 50 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.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 100 µL Conjugate
    9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    10.   Aspirate and wash 4 times.

    11. 100 µL Substrate Solution
    12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    13. 100 µL Stop Solution
    14.   Add 100 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. Adipose progenitor cell secretion of GM-CSF and MMP9 promotes a stromal and immunological microenvironment that supports breast cancer progression
      Authors: F Reggiani, V Labanca, P Mancuso, C Rabascio, G Talarico, S Orecchioni, A Manconi, F Bertolini
      Cancer Res., 2017;0(0):.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    2. Fungal immunomodulatory protein-fve could modulate airway remodel through by affect IL17 cytokine
      Authors: YT Lee, CT Wu, HL Sun, JL Ko, KH Lue
      J Microbiol Immunol Infect, 2017;0(0):.
      Species: Mouse
      Sample Type: BALF
    3. Carvedilol attenuates paraquat-induced lung injury by inhibition of proinflammatory cytokines, chemokine MCP-1, NF-?B activation and oxidative stress mediators
      Cytokine, 2016;88(0):144-153.
      Species: Mouse
      Sample Type: Plasma
    4. SHIP1-expressing mesenchymal stem cells regulate hematopoietic stem cell homeostasis and lineage commitment during aging.
      Authors: Iyer S, Brooks R, Gumbleton M, Kerr W
      Stem Cells Dev, 2015;24(9):1073-81.
      Species: Mouse
      Sample Type: Serum
    5. beta-Glucuronidase, a Regulator of Lyme Arthritis Severity, Modulates Lysosomal Trafficking and MMP-9 Secretion in Response to Inflammatory Stimuli.
      Authors: Bramwell K, Mock K, Ma Y, Weis J, Teuscher C, Weis J
      J Immunol, 2015;195(4):1647-56.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    6. Pharmacologic inhibition of S-nitrosoglutathione reductase protects against experimental asthma in BALB/c mice through attenuation of both bronchoconstriction and inflammation.
      Authors: Blonder J, Mutka S, Sun X, Qiu J, Green L, Mehra N, Boyanapalli R, Suniga M, Look K, Delany C, Richards J, Looker D, Scoggin C, Rosenthal G
      BMC Pulm Med, 2014;14(0):3.
      Species: Mouse
      Sample Type: Plasma
    7. Activation of transcription factor Nrf2 signalling by the sphingosine kinase inhibitor SKI-II is mediated by the formation of Keap1 dimers.
      Authors: Mercado, Nicolas, Kizawa, Yasuo, Ueda, Keitaro, Xiong, Yeping, Kimura, Genki, Moses, Audric, Curtis, Jonathan, Ito, Kazuhiro, Barnes, Peter J
      PLoS ONE, 2014;9(2):e88168.
      Species: Mouse
      Sample Type: BALF
    8. A novel molecule Me6TREN promotes angiogenesis via enhancing endothelial progenitor cell mobilization and recruitment.
      Authors: Chen H, Wang S, Zhang J, Ren X, Zhang R, Shi W, Lv Y, Zhou Y, Yan X, Chen L, He L, Zhang B, Nan X, Yue W, Li Y, Pei X
      Sci Rep, 2014;4(0):6222.
    9. Eosinophils promote generation and maintenance of immunoglobulin-A-expressing plasma cells and contribute to gut immune homeostasis.
      Authors: Chu V, Beller A, Rausch S, Strandmark J, Zanker M, Arbach O, Kruglov A, Berek C
      Immunity, 2014;40(4):582-93.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    10. 12-Hydroxyheptadecatrienoic acid promotes epidermal wound healing by accelerating keratinocyte migration via the BLT2 receptor.
      Authors: Liu M, Saeki K, Matsunobu T, Okuno T, Koga T, Sugimoto Y, Yokoyama C, Nakamizo S, Kabashima K, Narumiya S, Shimizu T, Yokomizo T
      J Exp Med, 2014;211(6):1063-78.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    11. Notch-1 mediated cardiac protection following embryonic and induced pluripotent stem cell transplantation in doxorubicin-induced heart failure.
      Authors: Merino H, Singla D
      PLoS ONE, 2014;9(7):e101024.
      Species: Mouse
      Sample Type: Tissue Homogenates
    12. Soluble E-cadherin: a critical oncogene modulating receptor tyrosine kinases, MAPK and PI3K/Akt/mTOR signaling.
      Authors: Brouxhon, S M, Kyrkanides, S, Teng, X, Athar, M, Ghazizadeh, S, Simon, M, O'Banion, M K, Ma, L
      Oncogene, 2014;33(2):225-35.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    13. The combination of the histone deacetylase inhibitor vorinostat and synthetic triterpenoids reduces tumorigenesis in mouse models of cancer.
      Authors: Tran, Kim, Risingsong, Renee, Royce, Darlene, Williams, Charlott, Sporn, Michael, Pioli, Patricia, Gediya, Lalji K, Njar, Vincent, Liby, Karen T
      Carcinogenesis, 2013;34(1):199-210.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    14. Tissue redox activity as a hallmark of carcinogenesis: from early to terminal stages of cancer.
      Authors: Bakalova R, Zhelev Z, Aoki I, Saga T
      Clin Cancer Res, 2013;19(9):2503-17.
      Species: Mouse
      Sample Type: Plasma
    15. A hypomorphic mutation in the Gfi1 transcriptional repressor results in a novel form of neutropenia.
      Eur J Immunol, 2012;42(9):2395-408.
      Species: Mouse
      Sample Type: BALF
    16. The synthetic triterpenoid CDDO-methyl ester delays estrogen receptor-negative mammary carcinogenesis in polyoma middle T mice.
      Authors: Tran K, Risingsong R, Royce D
      Cancer Prev Res (Phila), 2012;5(5):726-34.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    17. The effects of low levels of dystrophin on mouse muscle function and pathology.
      Authors: van Putten M, Hulsker M, Nadarajah VD, van Heiningen SH, van Huizen E, van Iterson M, Admiraal P, Messemaker T, den Dunnen JT, 't Hoen PA, Aartsma-Rus A
      PLoS ONE, 2012;7(2):e31937.
      Species: Mouse
      Sample Type: Serum
    18. Role of c-Jun N-terminal protein kinase 1/2 (JNK1/2) in macrophage-mediated MMP-9 production in response to Moraxella catarrhalis lipooligosaccharide (LOS).
      Authors: Hassan F, Ren D, Zhang W
      PLoS ONE, 2012;7(5):e37912.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    19. Methamphetamine-associated cleavage of the synaptic adhesion molecule intercellular adhesion molecule-5.
      Authors: Conant K, Lonskaya I, Szklarczyk A, Krall C, Steiner J, Maguire-Zeiss K, Lim ST
      J. Neurochem., 2011;118(4):521-32.
      Species: Mouse
      Sample Type: Cell Lysates
    20. Activation of matrix metalloproteinases following anti-Abeta immunotherapy; implications for microhemorrhage occurrence.
      Authors: Wilcock DM, Morgan D, Gordon MN, Taylor TL, Ridnour LA, Wink DA, Colton CA
      J Neuroinflammation, 2011;8(0):115.
      Species: Mouse
      Sample Type: Tissue Homogenates
    21. Early protection from burn-induced acute lung injury by deletion of preprotachykinin-A gene.
      Authors: Sio SW, Moochhala S, Lu J
      Am. J. Respir. Crit. Care Med., 2010;181(1):36-46.
      Species: Mouse
      Sample Type: Tissue Homogenates
    22. Inhibition of EGFR pathway signaling and the metastatic potential of breast cancer cells by PA-MSHA mediated by type 1 fimbriae via a mannose-dependent manner.
      Authors: Liu ZB, Hou YF, Zhu J
      Oncogene, 2010;29(20):2996-3009.
      Species: Mouse
      Sample Type: Serum
    23. Galectin-3 contributes to neonatal hypoxic-ischemic brain injury.
      Authors: Doverhag C, Hedtjarn M, Poirier F, Mallard C, Hagberg H, Karlsson A, Savman K
      Neurobiol. Dis., 2010;38(1):36-46.
      Species: Mouse
      Sample Type: Tissue Homogenates
    24. 15-Epi-lipoxin A(4) inhibits the progression of endometriosis in a murine model.
      Authors: Chen QH, Zhou WD, Pu DM, Huang QS, Li T, Chen QX
      Fertil. Steril., 2010;93(5):1440-7.
      Species: Mouse
      Sample Type: Peritoneal Fluid
    25. Attenuation of AD-like neuropathology by harnessing peripheral immune cells: local elevation of IL-10 and MMP-9.
      Authors: Koronyo-Hamaoui M, Ko MK, Koronyo Y, Azoulay D, Seksenyan A, Kunis G, Pham M, Bakhsheshian J, Rogeri P, Black KL, Farkas DL, Schwartz M
      J. Neurochem., 2009;111(6):1409-24.
      Species: Mouse
      Sample Type: Tissue Homogenates
    26. Inducible nitric oxide synthase deficiency impairs matrix metalloproteinase-9 activity and disrupts leukocyte migration in hepatic ischemia/reperfusion injury.
      Authors: Hamada T, Duarte S, Tsuchihashi S, Busuttil RW, Coito AJ
      Am. J. Pathol., 2009;174(6):2265-77.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    27. Matrix metalloproteinase 9 facilitates West Nile virus entry into the brain.
      Authors: Wang P, Dai J, Bai F, Kong KF, Wong SJ, Montgomery RR, Madri JA, Fikrig E
      J. Virol., 2008;82(18):8978-85.
      Species: Mouse
      Sample Type: Serum
    Expand to show all 27 Citations
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