Total MMP-2 Quantikine ELISA Kit

For human, mouse, rat, porcine, and canine samples.
(22 citations)   
  • 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), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    0.082 ng/mL
  • Assay Range
    0.5 - 32 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Recombinant MMP-2 and natural human, mouse, rat, porcine, and canine active, pro-, and TIMP complexed MMP-2.
  • 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.
Control Available
QC126, Quantikine Immunoassay Control Set 923 for Total MMP-2 - Please Inquire
Product Summary
The Quantikine Total MMP-2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure MMP-2 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse/rat pro-MMP-2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural MMP-2 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring MMP-2.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean4.311.222.84.2611.121.5
Standard Deviation0.3020.420.8280.2750.7381.51
CV%73.83.66.56.67

Recovery

The recovery of Total MMP-2 spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 99 90-105
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Total MMP-2 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Total MMP-2 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: MMP-2

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 2
    • Entrez Gene IDs
      4313 (Human); 17390 (Mouse);
    • Alternate Names
      72 kDa gelatinase; CLG4; CLG4A72 kDa type IV collagenase; collagenase type IV-A; EC 3.4.24; EC 3.4.24.24; Gelatinase A; matrix metallopeptidase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IVcollagenase); matrix metalloproteinase 2 (gelatinase A, 72kD gelatinase, 72kD type IVcollagenase); Matrix metalloproteinase-2; matrix metalloproteinase-II; MMP2; MMP-2; MMP-II; MONA; neutrophil gelatinase; TBE-1matrix metalloproteinase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IVcollagenase);
    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 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.

    22 Citations: Showing 1 - 10
    Filter your results:

    Species
    Sample Type
    1. Aurora Kinase A is a Biomarker for Bladder Cancer Detection and Contributes to its Aggressive Behavior
      Authors: A Mobley, S Zhang, J Bondaruk, Y Wang, T Majewski, NP Caraway, L Huang, E Shoshan, G Velazquez-, G Nitti, S Lee, JG Lee, E Fuentes-Ma, D Willis, L Zhang, CC Guo, H Yao, K Baggerly, Y Lotan, SP Lerner, C Dinney, D McConkey, M Bar-Eli, B Czerniak
      Sci Rep, 2017;7(0):40714.
      Species: Human
      Sample Type: Cell Culture Supernates
    2. Comparison of fibrin glue and Vicryl sutures in conjunctival autografting for pterygium surgery
      Authors: X Wang, Y Zhang, L Zhou, R Wei, L Dong
      Mol. Vis., 2017;23(0):275-285.
      Species: Human
      Sample Type: Tears
    3. TIMP1 and MMP9 are predictors of mortality in septic patients in the emergency department and intensive care unit unlike MMP9/TIMP1 ratio: Multivariate model
      Authors: ME Niño, SE Serrano, DC Niño, DM McCosham, ME Cardenas, VP Villareal, M Lopez, A Pazin-Filh, FA Jaimes, F Cunha, R Schulz, D Torres-Due
      PLoS ONE, 2017;12(2):e0171191.
      Species: Human
      Sample Type: Serum
    4. Beneficial Effects of Galectin-3 Blockade in Vascular and Aortic Valve Alterations in an Experimental Pressure Overload Model
      Authors: J Ibarrola, E Martínez-M, JR Sádaba, V Arrieta, A García-Peñ, V Álvarez, A Fernández-, A Gainza, P Rossignol, V Cachofeiro, N López-Andr
      Int J Mol Sci, 2017;18(8):.
      Species: Rat
      Sample Type: Tissue Homogenates
    5. Differential diagnosis of lung cancer, its metastasis and chronic obstructive pulmonary disease based on serum Vegf, Il-8 and MMP-9
      Sci Rep, 2016;6(0):36065.
      Species: Human
      Sample Type: Serum
    6. MMP-2 Isoforms in Aortic Tissue and Serum of Patients with Ascending Aortic Aneurysms and Aortic Root Aneurysms
      PLoS ONE, 2016;11(11):e0164308.
      Species: Human
      Sample Type: Serum
    7. 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 Type: Serum
    8. Magnobovatol inhibits smooth muscle cell migration by suppressing PDGF-R? phosphorylation and inhibiting matrix metalloproteinase-2 expression
      Authors: H Kang, DH Ahn, JH Pak, KH Seo, NI Baek, SW Jang
      Int J Mol Med, 2016;37(5):1239-46.
      Species: Human
      Sample Type: Cell Culture Supernates
    9. CP-25, a novel compound, protects against autoimmune arthritis by modulating immune mediators of inflammation and bone damage
      Sci Rep, 2016;6(0):26239.
      Species: Rat
      Sample Type: Synovial Fluid
    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 Type: Cell Lysates
    11. 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 Type: Cell Culture Supernates
    12. Aqueous and alcoholic extracts of Triphala and their active compounds chebulagic acid and chebulinic acid prevented epithelial to mesenchymal transition in retinal pigment epithelial cells, by inhibiting SMAD-3 phosphorylation.
      Authors: Sivasankar S, Lavanya R, Brindha P, Angayarkanni N
      PLoS ONE, 2015;10(3):e0120512.
      Species: Human
      Sample Type: Cell Culture Supernates
    13. Oxidative and pro-inflammatory impact of regular and denicotinized cigarettes on blood brain barrier endothelial cells: is smoking reduced or nicotine-free products really safe?
      Authors: Naik, Pooja, Fofaria, Neel, Prasad, Shikha, Sajja, Ravi K, Weksler, Babette, Couraud, Pierre-O, Romero, Ignacio, Cucullo, Luca
      BMC Neurosci, 2014;15(0):51.
      Species: Human
      Sample Type: Cell Culture Supernates
    14. Role of serum VEGFA, TIMP2, MMP2 and MMP9 in monitoring response to adjuvant radiochemotherapy in patients with primary cervical cancer--results of a companion protocol of the randomized NOGGO-AGO phase III clinical trial.
      Authors: Braicu E, Gasimli K, Richter R, Nassir M, Kummel S, Blohmer J, Yalcinkaya I, Chekerov R, Ignat I, Ionescu A, Mentze M, Fotopoulou C, Pop C, Lichtenegger W, Sehouli J
      Anticancer Res, 2014;34(1):385-91.
      Species: Human
      Sample Type: Serum
    15. WNT5A induces release of exosomes containing pro-angiogenic and immunosuppressive factors from malignant melanoma cells.
      Authors: Ekstrom E, Bergenfelz C, von Bulow V, Serifler F, Carlemalm E, Jonsson G, Andersson T, Leandersson K
      Mol Cancer, 2014;13(0):88.
      Species: Human
      Sample Type: Cell Culture Supernates
    16. Overexpression of EMMPRIN isoform 2 is associated with head and neck cancer metastasis.
      Authors: Huang Z, Tan N, Guo W, Wang L, Li H, Zhang T, Liu X, Xu Q, Li J, Guo Z
      PLoS ONE, 2014;9(4):e91596.
      Species: Human
      Sample Type: Cell Culture Supernates
    17. Effect of mesenchymal stem cells and extracts derived from the placenta on trophoblast invasion and immune responses.
      Authors: Choi, Jong Ho, Jung, Jieun, Na, Kyu-Hwan, Cho, Kyung Ji, Yoon, Tae Ki, Kim, Gi Jin
      Stem Cells Dev, 2014;23(2):132-45.
      Species: Human
      Sample Type: Cell Culture Supernates
    18. 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
    19. G protein-coupled receptor kinase 6 deficiency promotes angiogenesis, tumor progression, and metastasis.
      Authors: Raghuwanshi S, Smith N, Rivers E, Thomas A, Sutton N, Hu Y, Mukhopadhyay S, Chen X, Leung T, Richardson R
      J Immunol, 2013;190(10):5329-36.
      Species: Mouse
      Sample Type: Tissue Homogenates
    20. Role of serum concentration of VEGFR1 and TIMP2 on clinical outcome in primary cervical cancer: results of a companion protocol of the randomized, NOGGO-AGO phase III adjuvant trial of simultaneous cisplatin-based radiochemotherapy vs. carboplatin and paclitaxel containing sequential radiotherapy.
      Authors: Braicu E, Fotopoulou C, Chekerov R, Richter R, Blohmer J, Kummel S, Stamatian F, Yalcinkaya I, Mentze M, Lichtenegger W, Sehouli J
      Cytokine, 2013;61(3):755-8.
      Species: Human
      Sample Type: Serum
    21. Neutrophil-derived matrix metalloproteinase-9 is a potent activator of trypsinogen in acinar cells in acute pancreatitis.
      Authors: Awla D, Abdulla A, Syk I, Jeppsson B, Regner S, Thorlacius H
      J. Leukoc. Biol., 2012;91(5):711-9.
      Species: Mouse
      Sample Type: Plasma
    22. Celastrol suppresses allergen-induced airway inflammation in a mouse allergic asthma model.
      Authors: Kim DY, Park JW, Jeoung D, Ro JY
      Eur. J. Pharmacol., 2009;612(1):98-105.
      Species: Mouse
      Sample Type: BALF
    Expand to show all 22 Citations
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    Quantikine Immunoassay Control Set 923 for Total MMP-2

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