Human Total MMP-1 DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
(Total MMP-1). The Reagent Diluent recommended may be suitable for most cell culture supernate, serum, and plasma samples. The Reagent Diluent selected for use can alter the performance of an immunoassay. Reagent Diluent optimization for samples with complex matrices such as serum and plasma, may improve their performance in this assay.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
Background: MMP-1
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).
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human Total MMP-1 DuoSet ELISA
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.
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Citations: Showing 1 - 10
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Single-Cell Multi-Dimensional data analysis reveals the role of ARL4C in driving rheumatoid arthritis progression and Macrophage polarization dynamics
Authors: Tang, N;Luo, X;Ding, Z;Shi, Y;Cao, X;Wu, S;
International immunopharmacology
Species: Human
Sample Types: Cell Culture Supernates
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Analgesic, Anti-Inflammatory, and Chondroprotective Activities of Siraitia grosvenorii Residual Extract
Authors: Lee, YM;Kim, DS;
International journal of molecular sciences
Species: Human
Sample Types: Cell Culture Supernates
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Glyoxal-derived advanced glycation end products (GO-AGEs) with UVB critically induce skin inflammaging: in vitro and in silico approaches
Authors: Sultana, R;Parveen, A;Kang, MC;Hong, SM;Kim, SY;
Scientific reports
Species: Human
Sample Types: Cell Culture Supernates
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Pro-Resolving Mediators in Rotator Cuff Disease: How Is the Bursa Involved?
Authors: Klatte-Schulz, F;Bormann, N;Bonell, A;Al-Michref, J;Nguyen, HL;Klöckner, P;Thiele, K;Moroder, P;Seifert, M;Sawitzki, B;Wildemann, B;Duda, GN;
Cells
Species: Human
Sample Types: Tissue Culture Supernates
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Human Probiotic Lactobacillus paracasei-Derived Extracellular Vesicles Improve Tumor Necrosis Factor-?-Induced Inflammatory Phenotypes in Human Skin
Authors: Lee, KS;Kim, Y;Lee, JH;Shon, S;Kim, A;Pham, AVQ;Kim, C;Kim, DH;Kim, YK;Cho, EG;
Cells
Species: Human
Sample Types: Cell Culture Supernates
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Effects of Bacterial Lysates and Metabolites on Collagen Homeostasis in TNF-?-Challenged Human Dermal Fibroblasts
Authors: Huuskonen, L;Anglenius, H;Ahonen, I;Tiihonen, K;
Microorganisms
Species: Human
Sample Types: Cell Culture Supernates
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Role of triggering receptor expressed on myeloid cells-1 in the mechanotransduction signaling pathways that link low shear stress with inflammation
Authors: M Liu, AN Panagopoul, UM Oguz, S Samant, CH Vasa, DK Agrawal, YS Chatzizisi
Scientific Reports, 2023-03-21;13(1):4656.
Species: Human
Sample Types: Cell Culture Supernates
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Co-culture models of endothelial cells, macrophages, and vascular smooth muscle cells for the study of the natural history of atherosclerosis
Authors: M Liu, S Samant, CH Vasa, RM Pedrigi, UM Oguz, S Ryu, T Wei, DR Anderson, DK Agrawal, YS Chatzizisi
PLoS ONE, 2023-01-20;18(1):e0280385.
Species: Human
Sample Types: Cell Culture Supernates
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Association of the Heart Rate Variability Response to Active Standing with the Severity of Calcific Aortic Valve Disease: Novel Insights of a Neurocardiovascular Pathology
Authors: J Rodríguez-, JM Torres-Are, N Ávila-Vanz, R Springall, R Bojalil, O Infante, C Lerma, JC Echeverría
Journal of Clinical Medicine, 2022-08-16;11(16):.
Species: Human
Sample Types: Serum
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Evaluation in a Cytokine Storm Model In Vivo of the Safety and Efficacy of Intravenous Administration of PRS CK STORM (Standardized Conditioned Medium Obtained by Coculture of Monocytes and Mesenchymal Stromal Cells)
Authors: JP Lapuente, G Gómez, J Marco-Brua, P Fernández, P Desportes, J Sanz, M García-Gil, F Bermejo, JV San Martín, A Algaba, JC De Gregori, D Lapuente, A De Gregori, B Lapuente, S Gómez, MLV Andrés, A Anel
Biomedicines, 2022-05-08;10(5):.
Species: Human
Sample Types: Cell Culture Supernates
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Non-Invasive Physical Plasma Generated by a Medical Argon Plasma Device Induces the Expression of Regenerative Factors in Human Gingival Keratinocytes, Fibroblasts, and Tissue Biopsies
Authors: B Eggers, MB Stope, J Marciniak, W Götz, A Mustea, J Deschner, M Nokhbehsai, FJ Kramer
Biomedicines, 2022-04-13;10(4):.
Species: Human
Sample Types: Cell Lysates
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Activation of the Innate Immune Checkpoint CLEC5A on Myeloid Cells in the Absence of Danger Signals Modulates Macrophages' Function but Does Not Trigger the Adaptive T Cell Immune Response
Authors: MJ Tosiek, K Groesser, A Pekcec, M Zwirek, G Murugesan, E Borges
Journal of Immunology Research, 2022-02-25;2022(0):9926305.
Species: Human
Sample Types: Cell Culture Supernates
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Protective effects of Quercus acuta Thunb. fruit extract against UVB-induced photoaging through ERK/AP-1 signaling modulation in human keratinocytes
Authors: JA Hong, D Bae, KN Oh, DR Oh, Y Kim, Y Kim, S Jeong Im, EJ Choi, SG Lee, M Kim, C Jeong, CY Choi
BMC complementary medicine and therapies, 2022-01-04;22(1):6.
Species: Human
Sample Types: Cell Culture Supernates
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Modulation by 17,20S(OH)2pD of Fibrosis-Related Mediators in Dermal Fibroblast Lines from Healthy Donors and from Patients with Systemic Sclerosis
Authors: ML Brown Lobb, AT Slominski, KA Hasty, S Zhang, DD Miller, W Li, TK Kim, Z Janjetovic, RC Tuckey, IO Scott, LK Myers, AE Postlethwa
International Journal of Molecular Sciences, 2021-12-29;23(1):.
Species: Human
Sample Types: Cell Culture Supernates
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MMPs and TIMPs levels are correlated with anthropometric parameters, blood pressure, and endothelial function in obesity
Authors: S Boumiza, K Chahed, Z Tabka, MP Jacob, X Norel, G Ozen
Scientific Reports, 2021-10-08;11(1):20052.
Species: Human
Sample Types: Plasma
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Classical and Non-classical Fibrosis Phenotypes Are Revealed by Lung and Cardiac Like Microvascular Tissues On-Chip
Authors: A Akinbote, V Beltran-Sa, M Cherubini, R Visone, C Hajal, D Cobanoglu, K Haase
Frontiers in Physiology, 2021-10-06;12(0):735915.
Species: Human
Sample Types: Cell Culture Supernates
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Nobiletin Decreases Inflammatory Mediator Expression in Tumor Necrosis Factor-Stimulated Human Periodontal Ligament Cells
Authors: Y Hosokawa, I Hosokawa, K Ozaki
Mediators of Inflammation, 2021-07-10;2021(0):5535844.
Species: Human
Sample Types: Cell Culture Supernates
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Olig2 regulates p53-mediated apoptosis, migration and invasion of melanoma cells
Authors: JE Lee, S Ahn, H Jeong, S An, CH Myung, JA Lee, SC Hong, YJ Kim, JY Kim, JH Ryu, M Noh, KT Nam, JS Hwang
Scientific Reports, 2021-04-08;11(1):7778.
Species: Human
Sample Types: Cell Culture Supernates
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The Polymethoxy Flavonoid Sudachitin Inhibits Interleukin-1&beta-Induced Inflammatory Mediator Production in Human Periodontal Ligament Cells
Authors: Y Hosokawa, I Hosokawa, K Ozaki, T Matsuo
BioMed Research International, 2021-01-28;2021(0):8826586.
Species: Human
Sample Types: Cell Culture Supernates
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Widespread regulation of gene expression by glucocorticoids in chondrocytes from patients with osteoarthritis as determined by RNA-Seq
Authors: A Pemmari, T Leppänen, M Hämäläinen, T Moilanen, K Vuolteenah, E Moilanen
Arthritis Res Ther, 2020-11-17;22(1):271.
Species: Human
Sample Types: Cell Culture Supernates
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Silencing miR-202-3p increases MMP-1 and promotes a brain invasive phenotype in metastatic breast cancer cells
Authors: R Harati, S Hafezi, A Mabondzo, A Tlili
PLoS ONE, 2020-10-01;15(10):e0239292.
Species: Human
Sample Types: Cell Culture Supernates
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Syringaresinol Inhibits UVA-Induced MMP-1 Expression by Suppression of MAPK/AP-1 Signaling in HaCaT Keratinocytes and Human Dermal Fibroblasts
Authors: JH Oh, YH Joo, F Karadeniz, J Ko, CS Kong
Int J Mol Sci, 2020-06-01;21(11):.
Species: Human
Sample Types: Cell Culture Supernates
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Synthetic Retinoid Seletinoid G Improves Skin Barrier Function through Wound Healing and Collagen Realignment in Human Skin Equivalents
Authors: ES Lee, Y Ahn, IH Bae, D Min, NH Park, W Jung, SH Kim, YD Hong, WS Park, CS Lee
Int J Mol Sci, 2020-04-30;21(9):.
Species: Human
Sample Types: Cell Culture Supernatants
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Anticatabolic and Anti-Inflammatory Effects of Myricetin 3-O-&beta-d-Galactopyranoside in UVA-Irradiated Dermal Cells via Repression of MAPK/AP-1 and Activation of TGF&beta/Smad
Authors: JH Oh, F Karadeniz, JI Lee, SY Park, Y Seo, CS Kong
Molecules, 2020-03-14;25(6):.
Species: Human
Sample Types: Cell Culture Supernates
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Anti-Photoaging Effect of Plant Extract Fermented with Lactobacillus buchneri on CCD-986sk Fibroblasts and HaCaT Keratinocytes
Authors: YM Kang, CH Hong, SH Kang, DS Seo, SO Kim, HY Lee, HJ Sim, HJ An
J Funct Biomater, 2020-01-09;11(1):.
Species: Human
Sample Types: Cell Culture Supernates
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Caffeic Acid Phenethyl Ester Inhibits UV-Induced MMP-1 Expression by Targeting Histone Acetyltransferases in Human Skin
Authors: EJ Shin, S Jo, HK Choi, S Choi, S Byun, TG Lim
Int J Mol Sci, 2019-06-22;20(12):.
Species: Human
Sample Types: Cell Culture Media
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Cosmeceutical activities of ethanol extract and its ethyl acetate fraction from coffee silverskin
Authors: SH Xuan, KS Lee, HJ Jeong, YM Park, JH Ha, SN Park
Biomater Res, 2019-01-14;23(0):2.
Species: Human
Sample Types: Cell Culture Supernates
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1,25(OH)2D3 and dexamethasone additively suppress synovial fibroblast activation by CCR6+ T helper memory cells and enhance the effect of tumor necrosis factor alpha blockade
Authors: W Dankers, C González-L, N Davelaar, PS Asmawidjaj, AMC Mus, JMW Hazes, EM Colin, E Lubberts
Arthritis Res. Ther., 2018-09-20;20(1):212.
Species: Human
Sample Types: Cell Culture Supernates
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Inhibitory Effect of Opuntia humifusa Fruit Water Extract on Solar Ultraviolet-Induced MMP-1 Expression
Authors: AR Han, TG Lim, YR Song, M Jang, YK Rhee, HD Hong, MH Kim, HJ Kim, CW Cho
Int J Mol Sci, 2018-08-24;19(9):.
Species: Human
Sample Types: Cell Culture Supernates
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Functionally distinct disease-associated fibroblast subsets in rheumatoid arthritis
Authors: F Mizoguchi, K Slowikowsk, K Wei, JL Marshall, DA Rao, SK Chang, HN Nguyen, EH Noss, JD Turner, BE Earp, PE Blazar, J Wright, BP Simmons, LT Donlin, GD Kalliolias, SM Goodman, VP Bykerk, LB Ivashkiv, JA Lederer, N Hacohen, PA Nigrovic, A Filer, CD Buckley, S Raychaudhu, MB Brenner
Nat Commun, 2018-02-23;9(1):789.
Species: Human
Sample Types: Cell Culture Supernates
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Role of MMP-1 (-519A/G, -1607 1G/2G), MMP-3 (Lys45Glu), MMP-7 (-181A/G), and MMP-12 (-82A/G) Variants and Plasma MMP Levels on Obesity-Related Phenotypes and Microvascular Reactivity in a Tunisian Population
Authors: S Boumiza, S Bchir, H Ben Nasr, A Abbassi, MP Jacob, X Norel, Z Tabka, K Chahed
Dis. Markers, 2017-11-26;2017(0):6198526.
Species: Human
Sample Types: Plasma
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Double-stranded RNA released from damaged articular chondrocytes promotes cartilage degeneration via Toll-like receptor 3-interleukin-33 pathway
Authors: C Li, K Chen, H Kang, Y Yan, K Liu, C Guo, J Qi, K Yang, F Wang, L Guo, C He, L Deng
Cell Death Dis, 2017-11-02;8(11):e3165.
Species: Human
Sample Types: Tissue Homogenates
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Saikosaponin A inhibits IL-1?-induced inflammatory mediators in human osteoarthritis chondrocytes by activating LXR?
Authors: H Gao, Y Song, D Li, W Feng, J Liu
Oncotarget, 2017-09-30;8(51):88941-88950.
Species: Human
Sample Types: Cell Culture Supernates
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Resveratrol-Enriched Rice Attenuates UVB-ROS-Induced Skin Aging via Downregulation of Inflammatory Cascades
Authors: L Subedi, TH Lee, HM Wahedi, SH Baek, SY Kim
Oxid Med Cell Longev, 2017-08-16;2017(0):8379539.
Species: Human
Sample Types: Cell Culture Supernates
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Autologous chondrocyte implantation-derived synovial fluids display distinct responder and non-responder proteomic profiles
Authors: CH Hulme, EL Wilson, MJ Peffers, S Roberts, DM Simpson, JB Richardson, P Gallacher, KT Wright
Arthritis Res. Ther., 2017-06-30;19(1):150.
Species: Human
Sample Types: Synovial Fluid
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MMP-1 Activation Contributes to Airway Smooth Muscle Growth and Asthma Severity
Authors: Shams-Un-Nisa Naveed
Am. J. Respir. Crit. Care Med, 2017-04-15;0(0):.
Species: Human
Sample Types: BALF
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Epigenetically-driven anatomical diversity of synovial fibroblasts guides joint-specific fibroblast functions
Authors: M Frank-Bert, M Trenkmann, K Klein, E Karouzakis, H Rehrauer, A Bratus, C Kolling, M Armaka, A Filer, BA Michel, RE Gay, CD Buckley, G Kollias, S Gay, C Ospelt
Nat Commun, 2017-03-23;8(0):14852.
Species: Human
Sample Types: Cell Culture Supernates
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The effect of lipoaspirates vs. dissected abdominal fat on breast cancer cells in vitro
Authors: F Almarzouqi, HO Rennekampf, JP Stromps, Z Alharbi, N Pallua
Eur. J. Med. Res, 2017-03-21;22(1):10.
Species: Human
Sample Types: Cell Lysates
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The Methoxyflavonoid Isosakuranetin Suppresses UV-B-Induced Matrix Metalloproteinase-1 Expression and Collagen Degradation Relevant for Skin Photoaging
Int J Mol Sci, 2016-09-01;17(9):.
Species: Human
Sample Types: Cell Culture Supernates
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Dimethyl fumarate protection against collagen II degradation.
Authors: Li Y, Tang J, Hu Y
Biochem Biophys Res Commun, 2014-10-08;454(2):257-61.
Species: Human
Sample Types: Cell Culture Supernates
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Topical N-acetylcysteine accelerates wound healing in vitro and in vivo via the PKC/Stat3 pathway.
Authors: Tsai M, Huang H, Hsu J, Lai Y, Hsiao Y, Lu F, Chang H
Int J Mol Sci, 2014-05-02;15(5):7563-78.
Species: Human
Sample Types: Cell Culture Supernates
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Decreased PGE(2) content reduces MMP-1 activity and consequently increases collagen density in human varicose vein.
Authors: Gomez I, Benyahia C, Louedec L, Leseche G, Jacob M, Longrois D, Norel X
PLoS ONE, 2014-02-05;9(2):e88021.
Species: Human
Sample Types: Tissue Culture Supernates
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Interleukin-17A+ cell counts are increased in systemic sclerosis skin and their number is inversely correlated with the extent of skin involvement.
Authors: Truchetet M, Brembilla N, Montanari E, Lonati P, Raschi E, Zeni S, Fontao L, Meroni P, Chizzolini C
Arthritis Rheum, 2013-05-01;65(5):1347-56.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2011-02-25;54(2):212-7.
Species: Human
Sample Types: Vitreous Humor
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