Phone:
+1 877 302 8632
Fax:
+1 888 205 9894 (Toll-free)
E-Mail:
orders@antibodies-online.com

MMP2 ELISA Kit

MMP2 Reactivity: Human Colorimetric Sandwich ELISA 3.5-800 ng/mL Cell Culture Supernatant, Plasma, Serum
Catalog No. ABIN1979609
  • Target See all MMP2 ELISA Kits
    MMP2 (Matrix Metalloproteinase 2 (MMP2))
    Reactivity
    • 9
    • 8
    • 4
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    Human
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Detection Range
    3.5-800 ng/mL
    Minimum Detection Limit
    3.5 ng/mL
    Application
    ELISA
    Purpose
    Human MMP-2 ELISA Kit for cell culture supernatants, heparin treated plasma, and serum samples. EDTA and Citrate are not recommended.
    Sample Type
    Plasma, Cell Culture Supernatant, Serum
    Analytical Method
    Quantitative
    Specificity
    This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, MMP-1, - 3, -7, -8, -10, -12, -13, PARC, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    Sensitivity
    3.500 pg/mL
    Characteristics
    • Strip plates and additional reagents allow for use in multiple experiments
    • Quantitative protein detection
    • Establishes normal range
    • The best products for confirmation of antibody array data
    Components
    • Pre-Coated 96-well Strip Microplate
    • Wash Buffer
    • Stop Solution
    • Assay Diluent(s)
    • Lyophilized Standard
    • Biotinylated Detection Antibody
    • Streptavidin-Conjugated HRP
    • TMB One-Step Substrate
    Material not included
    • Distilled or deionized water
    • Precision pipettes to deliver 2 μL to 1 μL volumes
    • Adjustable 1-25 μL pipettes for reagent preparation
    • 100 μL and 1 liter graduated cylinders
    • Tubes to prepare standard and sample dilutions
    • Absorbent paper
    • Microplate reader capable of measuring absorbance at 450nm
    • Log-log graph paper or computer and software for ELISA data analysis
    Featured
    Discover our best selling MMP2 ELISA Kit
    Top Product
    Discover our top product MMP2 ELISA Kit
  • Application Notes
    Recommended Dilution for serum and plasma samples5 - 50 fold
    Sample Volume
    100 μL
    Plate
    Pre-coated
    Protocol
    1. Prepare all reagents, samples and standards as instructed in the manual.
    2. Add 100 μL of standard or sample to each well.
    3. Incubate 2.5 h at RT or O/N at 4 °C.
    4. Add 100 μL of prepared biotin antibody to each well.
    5. Incubate 1 h at RT.
    6. Add 100 μL of prepared Streptavidin solution to each well.
    7. Incubate 45 min at RT.
    8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 μL of Stop Solution to each well.
    11. Read at 450 nm immediately.
    Reagent Preparation
    1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
      2. Sample dilution: If your samples need to be diluted, 1x Assay Diluent (Item E) should be used for dilution of serum/plasma/culture supernatants/urine. Suggested dilution for normal serum/plasma: 5-50 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
      3. Assay Diluent (Item E) should be diluted 5-fold with deionized or distilled water before use.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL 1x Assay Diluent (Item E, Assay Diluent should be diluted 5-fold with deionized or distilled water before use) into Item C vial to prepare a 2 myg/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 180 µL MMP-2 standard from the vial of tem C, into a tube with 270 µL 1x Assay Diluent to prepare a 800 ng/mL stock standard solution. Pipette 300myl 1x Assay Diluent into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Assay Diluent serves as the zero standard (0 ng/mL). 200 µL 180 µL standard + 270 µL 200myl 200 µL 200 µL 200 µL 200 µL 800 320 128 51.2 20.5 8.2 3.3 0 ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL
      5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
      6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent and used in step 4 of Part VI Assay Procedure.
      7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 440-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 25 µL of HRP-Streptavidin concentrate into a tube with 11 ml 1x Assay Diluent to prepare a final 440 fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
    Assay Procedure
    1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
      2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
      3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
      4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
      5. Discard the solution. Repeat the wash as in step
      6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
      7. Discard the solution. Repeat the wash as in step
      8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
      9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
    Calculation of Results

    Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
    Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent Human MMP-2 concentration (ng/mL) O D =4 50 n m 0.001 0.01 0.1 1 10 0.1 10 100 1,000
    Sensitivity: The minimum detectable dose of MMP-2 is typically less than 3.5 ng/mL.
    Recovery: Recovery was determined by spiking various levels of human MMP-2 into human serum, plasma (Heparin) and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 81.27 70-93 Plasma 84.53 72-95 Cell culture media 77.25 69-92
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 93 94 92 Range ( %) 81-103 82-105 82-103 1:4 Average % of Expected 95 92 94 Range ( %) 82-104 80-103 84-104
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    Assay Precision
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Restrictions
    For Research Use only
  • Handling Advice
    Avoid repeated freeze-thaw cycles.
    Storage
    -20 °C
    Storage Comment
    The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
    Expiry Date
    6 months
  • Ağardan, Değim, Yılmaz, Altıntaş, Topal: "The Effectiveness of Raloxifene-Loaded Liposomes and Cochleates in Breast Cancer Therapy." in: AAPS PharmSciTech, Vol. 17, Issue 4, pp. 968-77, (2016) (PubMed).

    Kapelouzou, Tsourelis, Kaklamanis, Degiannis, Kogerakis, Cokkinos: "Serum and tissue biomarkers in aortic stenosis." in: Global cardiology science & practice, Vol. 2015, Issue 4, pp. 49, (2016) (PubMed).

    Slivka, Dearth, Keane, Meng, Medberry, Riggio, Reing, Badylak: "Fractionation of an ECM hydrogel into structural and soluble components reveals distinctive roles in regulating macrophage behavior." in: Biomaterials science, Vol. 2, Issue 10, pp. 1521-34, (2016) (PubMed).

    Domienik-Kar?owicz, Rymarczyk, Dzikowska-Diduch, Lisik, Chmura, Demkow, Pruszczyk: "Emerging markers of atherosclerosis before and after bariatric surgery." in: Obesity surgery, Vol. 25, Issue 3, pp. 486-93, (2015) (PubMed).

    Wang, Zhou, Lu, Zhang, Huang, Su: "Glycyrrhetinic acid potently suppresses breast cancer invasion and metastasis by impairing the p38 MAPK-AP1 signaling axis." in: Expert opinion on therapeutic targets, Vol. 19, Issue 5, pp. 577-87, (2015) (PubMed).

    Garratt, Sutanto, Ling, Looi, Iosifidis, Martinovich, Shaw, Kicic-Starcevich, Knight, Ranganathan, Stick, Kicic: "Matrix metalloproteinase activation by free neutrophil elastase contributes to bronchiectasis progression in early cystic fibrosis." in: The European respiratory journal, Vol. 46, Issue 2, pp. 384-94, (2015) (PubMed).

    Wang, Chang, Sun, Qian, Liu, Pei, Guo, Liu: "Angiotensin II Induces an Increase in Matrix Metalloproteinase 2 Expression in Aortic Smooth Muscle Cells of Ascending Thoracic Aortic Aneurysms Through JNK, ERK1/2, and p38 MAPK Activation." in: Journal of cardiovascular pharmacology, Vol. 66, Issue 3, pp. 285-93, (2015) (PubMed).

    Abdel-Latif: "Plasma Levels of Matrix Metalloproteinase (MMP)-2, MMP-9 and Tumor Necrosis Factor-α in Chronic Hepatitis C Virus Patients." in: The open microbiology journal, Vol. 9, pp. 136-40, (2015) (PubMed).

    Kiliç, Uçar, Özdemir, Colak, Bal, Ertu?rul: "Circulating matrix metalloproteinases and tissue inhibitors of metalloproteinases levels in pediatric patients with congenital heart disease: Relationship to cardiac functions." in: Anadolu kardiyoloji dergisi : AKD = the Anatolian journal of cardiology, Vol. 14, Issue 6, pp. 531-41, (2014) (PubMed).

    Gluba-Brzózka, Michalska-Kasiczak, Franczyk-Skóra, Nocu?, Banach, Rysz: "Markers of increased cardiovascular risk in patients with chronic kidney disease." in: Lipids in health and disease, Vol. 13, pp. 135, (2014) (PubMed).

    Yang, Wang, Wang, Xu, He, Wen, Yan, Su, Zhu: "Toll-like receptor 4 prompts human breast cancer cells invasiveness via lipopolysaccharide stimulation and is overexpressed in patients with lymph node metastasis." in: PLoS ONE, Vol. 9, Issue 10, pp. e109980, (2014) (PubMed).

    Sun, Wang, Zhou, Lu, Zhang, Chen, Zhao, Su: "Inhibitory effect of emodin on migration, invasion and metastasis of human breast cancer MDA-MB-231 cells in vitro and in vivo." in: Oncology reports, Vol. 33, Issue 1, pp. 338-46, (2014) (PubMed).

    Wang, Zhou, Du, Zhang, Lu, Su: "Baicalin suppresses migration, invasion and metastasis of breast cancer via p38MAPK signaling pathway." in: Anti-cancer agents in medicinal chemistry, Vol. 13, Issue 6, pp. 923-31, (2014) (PubMed).

    Hampel, Klonisch, Sel, Schulze, Garreis, Seitmann, Zouboulis, Paulsen: "Insulin-like factor 3 promotes wound healing at the ocular surface." in: Endocrinology, Vol. 154, Issue 6, pp. 2034-45, (2013) (PubMed).

    Yücel, De?im, Yilmaz: "Nanoparticle and liposome formulations of doxycycline: transport properties through Caco-2 cell line and effects on matrix metalloproteinase secretion." in: Biomedicine & pharmacotherapy, Vol. 67, Issue 6, pp. 459-67, (2013) (PubMed).

    Wang, Du, Zhang, Zhou, Lu, Zhang, Su: "Inhibitory Effects of PC-SPESII Herbal Extract on Human Breast Cancer Metastasis." in: Evidence-based complementary and alternative medicine : eCAM, Vol. 2013, pp. 894386, (2013) (PubMed).

    Chim, Armijo, Miller, Gliniak, Serret, Gosain: "Propranolol induces regression of hemangioma cells through HIF-1?-mediated inhibition of VEGF-A." in: Annals of surgery, Vol. 256, Issue 1, pp. 146-56, (2012) (PubMed).

    Toegel, Wu, Otero, Goldring, Leelapornpisid, Chiari, Kolb, Unger, Windhager, Viernstein: "Caesalpinia sappan extract inhibits IL1?-mediated overexpression of matrix metalloproteinases in human chondrocytes." in: Genes & nutrition, Vol. 7, Issue 2, pp. 307-18, (2012) (PubMed).

    Prasadam, Crawford, Xiao et al.: "Aggravation of ADAMTS and matrix metalloproteinase production and role of ERK1/2 pathway in the interaction of osteoarthritic subchondral bone osteoblasts and articular cartilage chondrocytes -- ..." in: The Journal of rheumatology, Vol. 39, Issue 3, pp. 621-34, (2012) (PubMed).

    Kaminska-Winciorek, Deja, Polanska, Jarosz-Chobot: "The role of selected metalloproteinases in cheiroarthropathy in children with type 1 diabetes - a pilotage study." in: International journal of clinical practice, Vol. 66, Issue 4, pp. 374-7, (2012) (PubMed).

  • Target See all MMP2 ELISA Kits
    MMP2 (Matrix Metalloproteinase 2 (MMP2))
    Alternative Name
    MMP-2 (MMP2 Products)
    Synonyms
    2-MMP ELISA Kit, CG1794 ELISA Kit, DM2-MMP ELISA Kit, Dm2-MMP ELISA Kit, Dmel\\CG1794 ELISA Kit, MMP2 ELISA Kit, anon-WO0118547.84 ELISA Kit, dm-2MMP ELISA Kit, dmmp2 ELISA Kit, l(2)02353 ELISA Kit, mmp2 ELISA Kit, wu:fa99h12 ELISA Kit, wu:fk89d01 ELISA Kit, fgmmp-2 ELISA Kit, Mmp-2 ELISA Kit, LOC100135793 ELISA Kit, Clg4a ELISA Kit, GelA ELISA Kit, MMP-2 ELISA Kit, CLG4 ELISA Kit, CLG4A ELISA Kit, MMP-II ELISA Kit, MONA ELISA Kit, TBE-1 ELISA Kit, matrix metallopeptidase 2 ELISA Kit, Matrix metalloproteinase 2 ELISA Kit, matrix metalloproteinase 2 ELISA Kit, matrix metalloproteinase-2 ELISA Kit, MMP2 ELISA Kit, Mmp2 ELISA Kit, mmp2 ELISA Kit, LOC657982 ELISA Kit, LOC100135793 ELISA Kit
    Background
    The Human MMP-2 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human MMP-2 in serum, plasma (Collect plasma using heparin as an anticoagulant. EDTA and Citrate are not recommended as anticoagulants for use in this assay due to their chelating properties), cell culture supernates and urine. This assay employs an antibody specific for human MMP-2 coated on a 96-well plate. Standards and samples are pipetted into the wells and MMP-2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human MMP-2 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of MMP-2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
    Gene ID
    4313
    UniProt
    P08253
    Pathways
    Activation of Innate immune Response
You are here:
Support