Human Total MMP-1 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY901B
DY901B-05
Ancillary Products Available
Human Total MMP-1 ELISA Standard Curve
2 Images
Product Details
Procedure
Citations (20)
FAQs
Supplemental Products
Reviews

Human Total MMP-1 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sufficient Materials
For five and fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human active and pro-Matrix Metalloproteinase 1
(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)

Data Examples

Human Total MMP-1 ELISA Standard Curve

Human Total MMP-1 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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).

Long Name:
Matrix Metalloproteinase 1
Entrez Gene IDs:
4312 (Human); 83995 (Mouse)
Alternate Names:
CLGmatrix metalloprotease 1; CLGN; EC 3.4.24; EC 3.4.24.7; Fibroblast collagenase; interstitial collagenase; matrix metallopeptidase 1 (interstitial collagenase); matrix metalloproteinase 1 (interstitial collagenase); Matrix metalloproteinase-1; MMP1; MMP-1

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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.

20 Citations: Showing 1 - 10
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  1. 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;21(9):.
    Species: Human
    Sample Types: Cell Culture Supernatants
  2. 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;11(1):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. 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;23(0):2.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. 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;19(9):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. 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;9(1):789.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. 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;20(1):212.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. 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;22(1):10.
    Species: Human
    Sample Types: Cell Lysates
  8. 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;8(0):14852.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. 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;2017(0):8379539.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. 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;19(1):150.
    Species: Human
    Sample Types: Synovial Fluid
  11. 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;8(51):88941-88950.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. 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;2017(0):6198526.
    Species: Human
    Sample Types: Plasma
  13. 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;8(11):e3165.
    Species: Human
    Sample Types: Tissue Homogenates
  14. MMP-1 Activation Contributes to Airway Smooth Muscle Growth and Asthma Severity
    Authors: Shams-Un-Nisa Naveed
    Am. J. Respir. Crit. Care Med, 2016;0(0):.
    Species: Human
    Sample Types: BALF
  15. The Methoxyflavonoid Isosakuranetin Suppresses UV-B-Induced Matrix Metalloproteinase-1 Expression and Collagen Degradation Relevant for Skin Photoaging
    Int J Mol Sci, 2016;17(9):.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. 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;9(2):e88021.
    Species: Human
    Sample Types: Tissue Culture Supernates
  17. Dimethyl fumarate protection against collagen II degradation.
    Authors: Li Y, Tang J, Hu Y
    Biochem Biophys Res Commun, 2014;454(2):257-61.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. 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;15(5):7563-78.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. 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;65(5):1347-56.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. 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 Types: Vitreous Humor

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