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Human MMP-9(Matrix Metalloproteinase 9) ELISA Kit

Citations(40)Uniprot : P14780

  • Cat.No.:E-EL-H6075

  • Reactivity: Human
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  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
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Product Details

Properties

Assay type Sandwich-ELISA
Format 96T/48T
Assay time 3.5h
Detection range 0.16-10 ng/mL
Sensitivity 0.10 ng/mL
Sample type &Sample volume serum, plasma and other biological fluids; 100μL
Specificity This kit recognizes Human MMP-9 in samples. No significant cross-reactivity or interference between Human MMP-9 and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of Human MMP-9 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human MMP-9. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human MMP-9 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human MMP-9, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human MMP-9. You can calculate the concentration of Human MMP-9 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
ItemSpecificationsStorage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips 		-20℃, 6 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL 		-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8°C
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human MMP-9 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human MMP-9 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 0.47 1.05 4.76 0.48 1.15 4.56
Standard deviation 0.03 0.05 0.20 0.03 0.06 0.19
CV (%) 6.38 4.76 4.20 6.25 5.22 4.17

Recovery

The recovery of Human MMP-9 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum(n=8) 87-97 93
EDTA plasma (n=8) 98-110 103
Cell culture media (n=8) 90-102 95

Linearity

Samples were spiked with high concentrations of Human MMP-9 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database LinksSwissProt:   P14780
SynonymsMMP9, CLG4B,Gelatinase B, GELB, MANDP2, 92kDa Type IV Collagenase, 92 KDa Gelatinase
Research AreaCancer, Cell Biology, Cardiovascular, Signal transduction

Assay Procedures

elisa assay procedure 1

1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
elisa assay procedure 2

2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
elisa assay procedure 3

3. Aspirate and wash the plate for 3 times
elisa assay procedure 4

4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
elisa assay procedure 5

5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
elisa assay procedure 6

6. Add 50μL Stop Solution
elisa assay procedure 7

7. Read the plate at 450nm immediately. Calculation of the results

Citations

  1. JOURNAL OF HEPATOLOGY (2019) IF: 18.946
    MMP2/MMP9-mediated CD100 shedding is crucial for inducing intrahepatic anti-HBV CD8 T cell responses and HBV clearance

    DOI: 10.1016/j.jhep.2019.05.013

    PMID: 31173811

    		 Sample: Serum
  2. Nature Communications (2021) IF: 14.919
    Geometrically encoded SERS nanobarcodes for the logical detection of nasopharyngeal carcinoma-related progression biomarkers

    DOI: 10.1038/s41467-021-23789-3

    PMID: 34078895

    		 Sample: plasma
  3. Science Signaling (2016) IF: 7.359
    Quantitative phosphoproteomic analysis identifies the critical role of JNK1 in neuroinflammation induced by Japanese encephalitis virus

    DOI: 10.1126/scisignal.aaf5132
    Sample: Human glioma cell line U251
  4. Marine Drugs (2023) IF: 6.085
    Mechanisms of Antitumor Invasion and Metastasis of the Marine Fungal Derivative Epi-Aszonalenin A in HT1080 Cells

    DOI: 10.3390/md21030156
    Sample: HT1080 cell
  5. Journal of Materials Chemistry B (2022) IF: 6.331
    Dual-Sensitive Fluorescent Nanoprobes for Detection of Matrix Metalloproteinases and Low pH in 3D Tumor Microenvironment

    DOI: 10.1039/D2TB00519K
    Sample: U2OS cell
  6. Cells (2021) IF: 6.6
    The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases

    DOI: 10.3390/cells10123506
    Sample: cardiac fibroblast
  7. Frontiers in Immunology (2020) IF: 5.085
    Infection of Mycobacterium tuberculosis Promotes Both M1/M2 Polarization and MMP Production in Cigarette Smoke-Exposed Macrophages

    DOI: 10.3389/fimmu.2020.01902

    PMID: 32973788

    		 Sample: Serum
  8. CHEMICO-BIOLOGICAL INTERACTIONS (2021) IF: 5.194
    A novel glyceroglycolipid from brown algae Ishige okamurae improve photoaging and counteract inflammation in UVB-induced HaCaT cells

    DOI: 10.1016/j.cbi.2021.109737

    PMID: 34740599

    		 Sample: HaCaT cells
  9. INTERNATIONAL JOURNAL OF ONCOLOGY (2014) IF: 5.65
    Downregulation of FoxM1 inhibits proliferation, invasion and angiogenesis of HeLa cells in vitro and in vivo

    DOI: 10.3892/ijo.2014.2645
    Sample: Cell culture supernatant
  10. FREE RADICAL BIOLOGY AND MEDICINE (2015) IF: 5.736
    Hypermethylation of the Keap1 gene inactivates its function, promotes Nrf2 nuclear accumulation, and is involved in arsenite-induced human keratinocyte transformation

    DOI: 10.1016/j.freeradbiomed.2015.07.153
    Sample: Cell culture supernatant,Cell lysate
  11. Frontiers in Bioscience-Landmark (2021) IF: 4.009
    Association between isotretinoin (Roaccutanne) use and changes in periodontal clinical parameters and MMP-8 and MMP-9 salivary levels.

    DOI: 10.52586/4933
    Sample: saliva
  12. American Journal of Translational Research (2021) IF: 4.06
    Effect of levetiracetam in combination with topiramate on immune function, cognitive function, and neuronal nutritional status of children with intractable epilepsy

    PMID: 34650715

    		 Sample: serum
  13. JOURNAL OF ETHNOPHARMACOLOGY (2021) IF: 4.36
    Anti-proliferation and anti-inflammation effects of corilagin in rheumatoid arthritis by downregulating NF-κB and MAPK signaling pathways

    DOI: 10.1016/j.jep.2021.114791

    PMID: 34737112

    		 Sample: MH7A cell
  14. MOLECULES (2022) IF: 4.412
    Salivary MMP-9 Levels in Chronic Periodontitis Patients with Type-II Diabetes Mellitus

    DOI: 10.3390/molecules27072174

    PMID: 35408573

    		 Sample: saliva
  15. MOLECULES (2019) IF: 3.06
    Ampelopsin E Reduces the Invasiveness of the Triple Negative Breast Cancer Cell Line, MDA-MB-231

    DOI: 10.3390/molecules24142619
    Sample: MDA-MB-231 cells
  16. INTEGRATIVE CANCER THERAPIES (2022) IF: 3.077
    Anti-metastatic Potential of Natural Triterpenoid Cucurbitacin B Against Cholangiocarcinoma Cells by Targeting Src Protein

    DOI: 10.1177/15347354221124861

    PMID: 36154723

    		 Sample: KKU-213 cell
  17. JOURNAL OF GENE MEDICINE (2020) IF: 3.258
    Human cytomegalovirus infection is correlated with atherosclerotic plaque vulnerability in carotid artery

    DOI: 10.1002/jgm.3236

    PMID: 32468600

    		 Sample: Serum
  18. International Journal of Environmental Research and Public Health (2021) IF: 3.39
    Salivary Metalloproteinase-8 and Metalloproteinase-9 Evaluation in Patients Undergoing Fixed Orthodontic Treatment before and after Periodontal Therapy

    DOI: 10.3390/ijerph18041583

    PMID: 33567492

    		 Sample: Saliva
  19. JOURNAL OF VASCULAR SURGERY (2020) IF: 3.405
    Magnetic resonance imaging assessment of proteolytic enzyme concentrations and biologic properties of intraluminal thrombus in abdominal aortic aneurysms

    DOI: 10.1016/j.jvs.2019.11.032
    Sample: aortic wall
  20. CLINICAL CHEMISTRY AND LABORATORY MEDICINE (2017) IF: 3.432
    Obesity, Metabolic syndrome, Diabetes

    DOI: 10.1515/cclm-2017-5030
    Sample: serum
  21. PLACENTA (2022) IF: 3.481
    Fenofibrate mediated activation of PPARα negatively regulates trophoblast invasion

    DOI: 10.1016/j.placenta.2022.05.015

    PMID: 35803128

    		 Sample: HTR8/SVneo cell
  22. Materials (2021) IF: 3.623
    Lutein Treatment Effects on the Redox Status and Metalloproteinase-9 (MMP-9) in Oral Cancer Squamous Cells—Are There Therapeutical Hopes?

    DOI: 10.3390/ma14112968
    Sample: Cell lysate
  23. Journal of Functional Foods (2020) IF: 3.701
    Trehalose against UVB-induced skin photoaging by suppressing MMP expression and enhancing procollagen I synthesis in HaCaT cells

    DOI: 10.1016/j.jff.2020.104198
    Sample: HaCaT cell
  24. TOXICOLOGY AND APPLIED PHARMACOLOGY (2015) IF: 3.705
    Continuous activation of Nrf2 and its target antioxidant enzymes leads to arsenite-induced malignant transformation of human bronchial epithelial cells

    DOI: 10.1016/j.taap.2015.09.020
    Sample: Cell culture supernatant
  25. CELLULAR SIGNALLING (2017) IF: 3.937
    Identification of NCK1 as a novel downstream effector of STAT3 in colorectal cancer metastasis and angiogenesis

    DOI: 10.1016/j.cellsig.2017.04.020
    Sample: Cell culture supernatant
  26. BMC Neurology (2021) IF: 2.474
    Association between serum NLRP3 and malignant brain edema in patients with acute ischemic stroke

    DOI: 10.1186/s12883-021-02369-4

    PMID: 34493232

    		 Sample: serum
  27. INTERNATIONAL JOURNAL OF CLINICAL PRACTICE (2021) IF: 2.503
    Plasma matrix metalloproteinase-9 level change with omalizumab treatment in chronic spontaneous urticaria

    DOI: 10.1111/ijcp.14942

    PMID: 34610203

    		 Sample: serum
  28. EUROPEAN ARCHIVES OF OTO-RHINO-LARYNGOLOGY (2021) IF: 2.503
    BMP-7, MMP-9, and TGF-β tissue remodeling proteins and their correlations with interleukins 6 and 10 in chronic rhinosinusitis

    DOI: 10.1007/s00405-021-06722-8

    PMID: 33715043

    		 Sample: Tissue homogenate
  29. Evidence-based Complementary and Alternative Medicine (2022) IF: 2.65
    The Predictive Values of MMP-9, PLTs, ESR, and CRP Levels in Kawasaki Disease with Cardiovascular Injury.

    DOI: 10.1155/2022/6913315
    Sample: serum
  30. Applied Sciences-Basel (2021) IF: 2.679
    Biochemical Mapping of the Inflamed Human Dental Pulp

    DOI: 10.3390/app112110395
    Sample: dental pulp lysates
  31. ANDROLOGIA (2022) IF: 2.775
    Relationship of interleukin-23 with matrix metalloproteinase-9, pentraxin-3, sleep quality and prostate size in benign prostatic hyperplasia

    DOI: 10.1111/and.14387

    PMID: 35092070

    		 Sample: serum
  32. ANNALS OF VASCULAR SURGERY (2022) IF: 1.607
    Correlation between proteolytic activity and abdominal aortic aneurysm wall morphology with intraluminal thrombus volume

    DOI: 10.1016/j.avsg.2022.05.039

    PMID: 35779804

    		 Sample: Abdominal Aortic Aneurysm (AAA) Wall
  33. BIOLOGIA (2022) IF: 1.653
    UPLC-ESI-QTOF-MS/MS and 1H-NMR identification of alkaloids in potent fraction of Catharanthus roseus leaves inhibits migration and invasion of MDA-MB-231 cells

    DOI: 10.1007/s11756-022-01185-6
    Sample: MDA-MB-231 cell
  34. EXPERIMENTAL AND CLINICAL ENDOCRINOLOGY & DIABETES (2017) IF: 1.685
    Expression and Significance of MMPs in Synovial Fluid, Serum and PBMC Culture Supernatant Stimulated by LPS in Osteoarthritis Patients With or Without Diabetes

    DOI: 10.1055/s-0043-122223

    PMID: 29270923

    		 Sample: Serum,Synovia
  35. Journal of Stroke & Cerebrovascular Diseases (2020) IF: 1.787
    Incremental Accuracy of Blood Biomarkers for Predicting Clinical Outcomes After Intracerebral Hemorrhage

    DOI: 10.1016/j.jstrokecerebrovasdis.2020.105537

    PMID: 33338706

    		 Sample: serum
  36. Acta Angiologica (2019)
    Monitoring of endostatin, TNF-a VEGFs, MMP-9, and cathepsin-L during three months of diosmin treatment in patients with chronic venous disease (CVD)

    DOI: 10.5603/AA.2019.0002
    Sample: plasma
  37. International Journal of Reproductive Biomedicine (2021)
    Matrix metalloproteinase-9 is elevated and related to interleukin-17 and psychological stress in male infertility: A cross-sectional study

    DOI: 10.18502/ijrm.v19i4.9059
    Sample: Spermatozoa (SPZ)
  38. Asian Pacific Journal of Cancer Prevention (2021)
    Cucurbitacin B Diminishes Metastatic Behavior of Cholangiocarcinoma Cells by Suppressing Focal Adhesion Kinase

    DOI: 10.31557/APJCP.2021.22.1.219

    PMID: 33507702

    		 Sample: KKU-452 cell
  39. SCIENTIA PHARMACEUTICA (2022)
    Colorectal Cancer Chemoprevention by S-Allyl Cysteine–Caffeic Acid Hybrids: In Vitro Biological Activity and In Silico Studies

    DOI: 10.3390/scipharm90030040
    Sample: SW480 cell
  40. BIOLOGIA (2020) IF: 0.8110
    Catharanthus roseus L. extract downregulates the expression profile of motility-related genes in highly invasive human breast cancer cell line MDA-MB-231

    DOI: 10.2478/s11756-020-00641-5
    Sample: MDA-MB-231 cell
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Reviews/Q&A

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Q

T*************rSubmitted [ Dec 24 2020 ]

Asked: Hello I used your Elabscience human MMP-9 kit for my study. But five journals rejected my article. Because the reference range of this kit is 0.16-10 ng / mL. In our study, we found normal values as 1.6-1.8. However, other studies show normal values as 20-30 ng/mL. This makes my work look questionable. How can I make a statement about this? I need to give an explanation to answer the questions. I will be glad if you help. Thanks in advance.

Reply

A

adminSubmitted [ Dec 24 2020 ]

Answered: Hi Tugba, Thanks for your message. Could you let us know which country you are from, so that I could forward to our local sales representative to contact you ASAP. Best regards

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