Human Total MMP-8 Quantikine ELISA Kit

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(20 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 (10 uL), Serum (10 uL), Heparin Plasma (10 uL), Saliva (10 uL)
  • Sensitivity
    0.06 ng/mL
  • Assay Range
    0.2 - 10 ng/mL (Cell Culture Supernates, Serum, Heparin Plasma, Saliva)
  • Specificity
    Natural and recombinant human MMP-8
  • 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
QC129, Quantikine Immunoassay Control Set 536 for Human Total MMP-8 - Please Inquire
Product Summary
The Quantikine Human Total MMP-8 Immunoassay is a 4.5 hour solid phase ELISA designed to measure total MMP-8 (pro- and active MMP-8) in cell culture supernates, serum, plasma, and saliva. It contains NS0-expressed recombinant human MMP-8 and antibodies raised against the recombinant protein. Natural human MMP-8 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-8.

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, Saliva
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean1.253.617.241.213.536.85
Standard Deviation0.0650.1820.3880.0730.1470.343
CV%5.255.464.25

Recovery

The recovery of MMP-8 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=8) 95 86-112
Heparin Plasma (n=5) 102 90-114
Serum (n=5) 97 88-105
Linearity
To assess the linearity of the assay, samples containing high concentrations of MMP-8 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Total MMP-8 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: MMP-8

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 8
    • Entrez Gene IDs
      4317 (Human); 17394 (Mouse); 63849 (Rat);
    • Alternate Names
      CLG1HNC; Collagenase 2; EC 3.4.24; EC 3.4.24.34; matrix metallopeptidase 8 (neutrophil collagenase); matrix metalloproteinase 8 (neutrophil collagenase); Matrix metalloproteinase-8; MMP8; MMP-8; neutrophil collagenase; PMNL collagenase; PMNL-CL;
    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. 150 µL Assay Diluent
    4.   Add 150 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.

    20 Citations: Showing 1 - 10
    Filter your results:

    Species
    Sample Type
    1. Identification of Gingival Crevicular Fluid Sampling, Analytical Methods, and Oral Biomarkers for the Diagnosis and Monitoring of Periodontal Diseases: A Systematic Review
      Authors: Z Nazar Maje, K Philip, AM Alabsi, S Pushparaja, D Swaminatha
      Dis. Markers, 2016;2016(0):1804727.
      Species: Human
      Sample Type: Saliva
    2. Relationship between Serum Levels of Metalloproteinase-8 and Tissue Inhibitor of Metalloproteinases-1 and Exercise Test Results in Postmenopausal Women
      Authors: J Mieczkowsk, E Rutkowska, J Mosiewicz, B Mosiewicz
      Dis. Markers, 2016;2016(0):7169531.
      Species: Human
      Sample Type: Serum
    3. The effect of type 2 diabetes mellitus and smoking on periodontal parameters and salivary matrix metalloproteinase-8 levels
      Authors: N Gupta, ND Gupta, S Garg, L Goyal, A Gupta, S Khan, S Moin
      J Oral Sci, 2016;58(1):1-6.
      Species: Human
      Sample Type: Saliva
    4. 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 Type: Subretinal Fluid
    5. 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 Type: Saliva
    6. Leptin administered in physiological or pharmacological doses does not regulate circulating angiogenesis factors in humans.
      Authors: Aronis KN, Diakopoulos KN, Fiorenza CG
      Diabetologia, 2011;54(9):2358-67.
      Species: Human
      Sample Type: Serum
    7. 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 Type: Vitreous Humor
    8. Increased levels of leukocyte-derived MMP-9 in patients with stable angina pectoris.
      Authors: Jonasson S, Lundberg A, Kalvegren H, Bergstrom I, Szymanowski A, Jonasson L
      PLoS ONE, 2011;6(4):e19340.
      Species: Human
      Sample Type: Serum
    9. Early Elevation of Matrix Metalloproteinase-8 and -9 in Pediatric ARDS Is Associated with an Increased Risk of Prolonged Mechanical Ventilation.
      Authors: Kong MY, Li Y, Oster R, Gaggar A, Clancy JP
      PLoS ONE, 2011;6(8):e22596.
      Species: Human
      Sample Type: Tissue Secretion
    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 Type: Cell Culture Supernates
    11. Serum matrix metalloproteinases in adult CF patients: Relation to pulmonary exacerbation.
      Authors: Roderfeld M, Rath T, Schulz R, Seeger W, Tschuschner A, Graf J, Roeb E
      J. Cyst. Fibros., 2009;8(5):338-47.
      Species: Human
      Sample Type: Serum
    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. Matrix metalloprotease-9 dysregulation in lower airway secretions of cystic fibrosis patients.
      Authors: Gaggar A, Li Y, Weathington N, Winkler M, Kong M, Jackson P, Blalock JE, Clancy JP
      Am. J. Physiol. Lung Cell Mol. Physiol., 2007;293(1):L96-L104.
      Species: Human
      Sample Type: Sputum
    14. Changes in periodontal and rheumatological conditions after 2 years in patients with juvenile idiopathic arthritis.
      Authors: Miranda LA, Braga F, Fischer RG, Sztajnbok FR, Figueredo CM, Gustafsson A
      J. Periodontol., 2006;77(10):1695-700.
      Species: Human
      Sample Type: Gingival Crevicular Fluid (GCF)
    15. Midtrimester amniotic fluid matrix metalloproteinase-8 (MMP-8) levels above the 90th percentile are a marker for subsequent preterm premature rupture of membranes.
      Authors: Biggio JR, Ramsey PS, Cliver SP, Lyon MD, Goldenberg RL, Wenstrom KD
      Am. J. Obstet. Gynecol., 2005;192(1):109-13.
      Species: Human
      Sample Type: Amniotic Fluid
    16. Aberrant neutrophil reactions in periodontitis.
      Authors: Figueredo CM, Fischer RG, Gustafsson A
      J. Periodontol., 2005;76(6):951-5.
      Species: Human
      Sample Type: Gingival Crevicular Fluid (GCF)
    17. 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
    18. Effect of tobacco smoking on neutrophil activity following periodontal surgery.
      Authors: Persson L, Bergstrom J, Gustafsson A
      J. Periodontol., 2003;74(10):1475-82.
      Species: Human
      Sample Type: Gingival Crevicular Fluid (GCF)
    19. 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 Type: Serum
    20. Shifts in biochemical markers associated with wound healing in laryngeal secretions following phonotrauma: a preliminary study.
      Authors: Verdolini K, Rosen CA, Branski RC, Hebda PA
      Ann. Otol. Rhinol. Laryngol., 2003;112(12):1021-5.
      Species: Human
      Sample Type: Tissue Secretion
    Expand to show all 20 Citations
    ELISA Controls
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    Quantikine Immunoassay Control Set 536 for Human Total MMP-8

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    Quantikine ELISA Wash Buffer 1

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    Average Rating: 5 (Based on 1 review)

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    Sample TestedCell culture supernatant

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