Human Decorin 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.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Decorin. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
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: Decorin
Decorin, also known as PG40 and PGS2, is a secreted chondroitin/dermatan sulfate proteoglycan in the family of small leucine-rich proteoglycans (SLRPs). Decorin regulates assembly of the extracellular collagen matrix and the bioactivity of the matrix associated growth factors basic FGF, GDF-8/Myostatin, TGF-beta, and WISP-1. It also binds and activates EGF R, ErbB4, and IGF-I R. Decorin promotes myoblast differentiation, supports angiogenesis, and inhibits tumor progression.
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 Decorin 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.
17
Citations: Showing 1 - 10
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Exploring the Impact of Astaxanthin Supplementation in Conjunction with a 12-Week CrossFit Training Regimen on Selected Adipo-Myokines Levels in Obese Males
Authors: Moqaddam, MA;Nemati, M;Dara, MM;Hoteit, M;Sadek, Z;Ramezani, A;Rand, MK;Abbassi-Daloii, A;Pashaei, Z;Almaqhawi, A;Razi, O;Escobar, KA;Supriya, R;Saeidi, A;Zouhal, H;
Nutrients
Species: Human
Sample Types: Plasma
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Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts
Authors: KEC Blokland, M Nizamoglu, H Habibie, T Borghuis, M Schuliga, BN Melgert, DA Knight, CA Brandsma, SD Pouwels, JK Burgess
Frontiers in Pharmacology, 2022-11-03;13(0):989169.
Species: Human
Sample Types: Cell Culture Supernates
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DCN released from ferroptotic cells ignites AGER-dependent immune responses
Authors: J Liu, S Zhu, L Zeng, J Li, DJ Klionsky, G Kroemer, J Jiang, D Tang, R Kang
Autophagy, 2021-12-29;0(0):1-14.
Species: Human
Sample Types: Cell Culture Supernates
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Nordic Walking Rather Than High Intensity Interval Training Reduced Myostatin Concentration More Effectively in Elderly Subjects and the Range of This Drop Was Modified by Metabolites of Vitamin D
Authors: K Micielska, M Flis, JA Kortas, E Rodziewicz, J Antosiewic, K Wochna, G Lombardi, E Ziemann
Nutrients, 2021-12-08;13(12):.
Species: Human
Sample Types: Serum
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Regulation of Cellular Senescence Is Independent from Profibrotic Fibroblast-Deposited ECM
Authors: KEC Blokland, H Habibie, T Borghuis, GJ Teitsma, M Schuliga, BN Melgert, DA Knight, CA Brandsma, SD Pouwels, JK Burgess
Cells, 2021-06-29;10(7):.
Species: Human
Sample Types: Cell Culture Supernates
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Exposed CendR Domain in Homing Peptide Yields Skin-Targeted Therapeutic in Epidermolysis Bullosa
Authors: T Pemmari, L Ivanova, U May, P Lingasamy, A Tobi, A Pasternack, S Prince, O Ritvos, S Makkapati, T Teesalu, MS Cairo, TAH Järvinen, Y Liao
Mol. Ther., 2020-05-20;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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Iron Status in Elderly Women Impacts Myostatin, Adiponectin and Osteocalcin Levels Induced by Nordic Walking Training
Authors: J Kortas, E Ziemann, D Juszczak, K Micielska, M Koz?owska, K Prusik, K Prusik, J Antosiewic
Nutrients, 2020-04-17;12(4):.
Species: Human
Sample Types: Plasma
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Anti-fibrotic effect of decorin in peritoneal dialysis and PD-associated peritonitis
Authors: N Jiang, Q Zhang, MK Chau, MS Yip, SL Lui, S Liu, KM Chu, HY Ngan, TM Chan, S Yung
EBioMedicine, 2020-02-12;52(0):102661.
Species: Human
Sample Types: Cell Culture Supernates, Cell Lysates
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Altered Decorin Biology in Proliferative Vitreoretinopathy: A Mechanistic and Cohort Study
Authors: G Begum, J O'Neill, R Chaudhary, K Blachford, DRJ Snead, M Berry, RAH Scott, A Logan, RJ Blanch
Invest. Ophthalmol. Vis. Sci., 2018-10-01;59(12):4929-4936.
Species: Human
Sample Types: Vitreous Humor
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Association of SLRPs with carotid artery atherosclerosis in essential hypertensive patients
Authors: Y Yang, QH Wu, Y Li, PJ Gao
J Hum Hypertens, 2018-06-05;0(0):.
Species: Human
Sample Types: Serum
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Surface functionalization of electrospun scaffolds using recombinant human decorin attracts circulating endothelial progenitor cells
Authors: S Hinderer, K Sudrow, M Schneider, M Holeiter, SL Layland, M Seifert, K Schenke-La
Sci Rep, 2018-01-08;8(1):110.
Species: Chinese Hamster
Sample Types: Cell Culture Supernates
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Higher Decorin Levels in Bone Marrow Plasma Are Associated with Superior Treatment Response to Novel Agent-Based Induction in Patients with Newly Diagnosed Myeloma - A Retrospective Study.
Authors: Huang S, Lin H, Yao M, Tang J, Wu S, Hou H, Chou W, Chou S, Hsu S, Ko B, Lu H, Tsay W, Tien H
PLoS ONE, 2015-09-17;10(9):e0137552.
Species: Human
Sample Types: Bone Marrow Fluid
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MicroRNA 181b regulates decorin production by dermal fibroblasts and may be a potential therapy for hypertrophic scar.
Authors: Kwan P, Ding J, Tredget E
PLoS ONE, 2015-04-02;10(4):e0123054.
Species: Human
Sample Types: Cell Culture Supernates
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Role of decorin in multiple myeloma (MM) bone marrow microenvironment.
Authors: Nemani N, Santo L, Eda H, Cirstea D, Mishima Y, Patel C, O'Donnell E, Yee A, Raje N
J Bone Miner Res, 2015-03-01;30(3):465-70.
Species: Human
Sample Types: Cell Culture Supernates
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Hepatocyte growth factor pathway upregulation in the bone marrow microenvironment in multiple myeloma is associated with lytic bone disease.
Authors: Kristensen I, Christensen J, Lyng M, Moller M, Pedersen L, Rasmussen L, Ditzel H, Abildgaard N
Br J Haematol, 2013-02-22;161(3):373-82.
Species: Human
Sample Types: Plasma
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Plasma resistin is associated with single nucleotide polymorphisms of a possible resistin receptor, the decorin gene, in the general Japanese population.
Authors: Onuma, Hiroshi, Tabara, Yasuharu, Kawamura, Ryoichi, Ohashi, Jun, Nishida, Wataru, Takata, Yasunori, Ochi, Masaaki, Nishimiya, Tatsuya, Kawamoto, Ryuichi, Kohara, Katsuhik, Miki, Tetsuro, Osawa, Haruhiko
Diabetes, 2012-11-27;62(2):649-52.
Species: Human
Sample Types: Plasma
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Plasma decorin predicts the presence of esophageal squamous cell carcinoma.
Authors: Wu IC, Wu DC, Huang CC
Int. J. Cancer, 2010-11-01;127(9):2138-46.
Species: Human
Sample Types: Plasma
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