Hyaluronan DuoSet ELISA

Catalog # Availability Size / Price Qty
DY3614-05
Ancillary Products Available
Hyaluronan ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (44)
FAQs
Supplemental Products
Reviews (2)

Hyaluronan DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
0.4 - 90 ng/mL
Sufficient Materials
For five 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 Hyaluronan. 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 reagent 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
  • Available in 5 (96-well) plate pack size
  • Economical alternative to complete kits

Kit Content

  • Capture Reagent
  • Detection Reagent
  • Natural Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 3 (5 plates): (Catalog # DY009) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 3.

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 # DY004), or 5% Tween 20 in PBS, 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

Hyaluronan ELISA Standard Curve

Product Datasheets

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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: Hyaluronan

Hyaluronan, also known as hyaluronic acid (HA) or sodium hyaluronate, is a naturally occurring linear polymer. Hyaluronan is a glycosaminoglycan (GAG) that is ubiquitously present in the extracellular matrix of all vertebrates and is also present in the capsule of some strains of Streptococci. Mammalian hyaluronan is synthesized by one of three distinct hyaluronan synthases (HAS1, 2, and 3), which produce HA polymers with different chain lengths and differ in their rates of synthesis. Whereas high molecular weight HA (> 500 kDa) is anti-angiogenic, anti-inflammatory and immunosuppressive, low molecular weight HA (10 - 500 kDa) is highly angiogenic and pro-inflammatory. HA oligomers are anti-apoptotic and upregulate heat shock protein expression. Functionally, hyaluronan molecules are important for the maintenance of a highly hydrated extracellular matrix in tissues, which is involved in cell adhesion and supports cell migration.

Alternate Names:
Hyaluronan

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 of 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 Hyaluronan 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.

44 Citations: Showing 1 - 10
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  1. Minimal Development of Liver Fibrosis in Adult Tolerant Liver Transplant Recipients Late After Immunosuppressive Drug Weaning and Transplantation
    Authors: AA Duizendstr, RJ De Knegt, NMA Nagtzaam, MGH Betjes, WA Dik, NHR Litjens, J Kwekkeboom
    Transplantation Proceedings, 2022;0(0):.
    Species: Human
    Sample Types: Serum
  2. UDP-glucose dehydrogenase (UGDH) activity is suppressed by peroxide and promoted by PDGF in fibroblast-like synoviocytes: Evidence of a redox control mechanism
    Authors: R Chandrasek, C Mathieu, R Sheth, AP Cheng, D Fong, R McCormack, H El-Gabalaw, S Alishetty, M Paige, CD Hoemann
    PLoS ONE, 2022;17(9):e0274420.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Synovial mesenchymal progenitor derived aggrecan regulates cartilage homeostasis and endogenous repair capacity
    Authors: RJ Krawetz, YE Wu, KL Bertram, A Shonak, AO Masson, G Ren, C Leonard, M Kapoor, JR Matyas, PT Salo
    Cell Death & Disease, 2022;13(5):470.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. No association between intravenous fluid volume and endothelial glycocalyx shedding in patients undergoing resuscitation for sepsis in the emergency department
    Authors: S Macdonald, E Bosio, NI Shapiro, L Balmer, S Burrows, M Hibbs, T Jowitt, L Smart, G Arendts, D Fatovich
    Scientific Reports, 2022;12(1):8733.
    Species: Human
    Sample Types: Serum
  5. SARS-CoV-2 RNA elements share human sequence identity and upregulate hyaluronan via NamiRNA-enhancer network
    Authors: W Li, S Yang, P Xu, D Zhang, Y Tong, L Chen, B Jia, A Li, C Lian, D Ru, B Zhang, M Liu, C Chen, W Fu, S Yuan, C Gu, L Wang, W Li, Y Liang, Z Yang, X Ren, S Wang, X Zhang, Y Song, Y Xie, H Lu, J Xu, H Wang, W Yu
    EBioMedicine, 2022;76(0):103861.
    Species: Human
    Sample Types: Plasma
  6. Alterations in plasma hyaluronic acid in patients with clinically stable COPD versus (non)smoking controls
    Authors: K Waeijen-Sm, NL Reynaert, RJHCG Beijers, S Houben-Wil, SO Simons, MA Spruit, FME Franssen
    Scientific Reports, 2021;11(1):15883.
    Species: Human
    Sample Types: Plasma
  7. A potent truncated form of human soluble CR1 is protective in a mouse model of renal ischemia-reperfusion injury
    Authors: AK Bongoni, IB Vikstrom, JL McRae, EJ Salvaris, N Fisicaro, MJ Pearse, S Wymann, T Rowe, AB Morelli, MP Hardy, PJ Cowan
    Scientific Reports, 2021;11(1):21873.
    Species: Mouse
    Sample Types: Serum
  8. Investigation of synovial fluid lubricants and inflammatory cytokines in the horse: a comparison of recombinant equine interleukin 1 beta-induced synovitis and joint lavage models
    Authors: A Watkins, D Fasanello, D Stefanovsk, S Schurer, K Caracappa, A D'Agostino, E Costello, H Freer, A Rollins, C Read, J Su, M Colville, M Paszek, B Wagner, H Reesink
    Bmc Veterinary Research, 2021;17(1):189.
    Species: Equine
    Sample Types: Synovial Fluid
  9. Role of FGF and Hyaluronan in Choroidal Neovascularization in Sorsby Fundus Dystrophy
    Authors: A Wolk, D Hatipoglu, A Cutler, M Ali, L Bell, J Hua Qi, R Singh, J Batoki, L Karle, VL Bonilha, O Wessely, H Stoehr, V Hascall, B Anand-Apte
    Cells, 2020;9(3):.
    Species: Human
    Sample Types: Plasma
  10. Loss of versican and production of hyaluronan in lung epithelial cells are associated with airway inflammation during RSV infection
    Authors: GG Kellar, KA Barrow, LM Rich, JS Debley, TN Wight, SF Ziegler, SR Reeves
    J Biol Chem, 2020;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  11. Characteristics of Hyaluronan Synthesis Inhibition by 4-Methylumbelliferone in Orbital Fibroblasts
    Authors: E Galgoczi, F Jeney, M Katko, A Erdei, A Gazdag, L Sira, M Bodor, E Berta, B Ujhelyi, Z Steiber, F Gyory, EV Nagy
    Invest. Ophthalmol. Vis. Sci., 2020;61(2):27.
    Species: Human
    Sample Types: Cell Cultue Supernates
  12. 4-Methylumbelliferone Inhibits Cancer Stem Cell Activation and Overcomes Chemoresistance in Ovarian Cancer
    Authors: NA Lokman, ZK Price, EK Hawkins, AM Macpherson, MK Oehler, C Ricciardel
    Cancers (Basel), 2019;11(8):.
    Species: Human
    Sample Types: Serum
  13. High levels of serum hyaluronan is an early predictor of dengue warning signs and perturbs vascular integrity
    Authors: CY Lin, C Kolliopoul, CH Huang, J Tenhunen, CH Heldin, YH Chen, P Heldin
    EBioMedicine, 2019;0(0):.
    Species: Human
    Sample Types: Serum
  14. Hyaluronan synthase 2 (HAS2) overexpression diminishes the procatabolic activity of chondrocytes by a mechanism independent of extracellular hyaluronan
    Authors: S Ishizuka, S Tsuchiya, Y Ohashi, K Terabe, EB Askew, N Ishizuka, CB Knudson, W Knudson
    J. Biol. Chem., 2019;0(0):.
    Species: Bovine
    Sample Types: Cell Culture Supernates
  15. Drug Repositioning Inferred from E2F1-Coregulator Interactions Studies for the Prevention and Treatment of Metastatic Cancers
    Authors: D Goody, SK Gupta, D Engelmann, A Spitschak, S Marquardt, S Mikkat, C Meier, C Hauser, JP Gundlach, JH Egberts, H Martin, T Schumacher, A Trauzold, O Wolkenhaue, S Logotheti, BM Pützer
    Theranostics, 2019;9(5):1490-1509.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Bioengineered tumor microenvironments with naked mole rats high-molecular-weight hyaluronan induces apoptosis in breast cancer cells
    Authors: Y Zhao, S Qiao, X Hou, H Tian, S Deng, K Ye, Y Nie, X Chen, H Yan, W Tian
    Oncogene, 2019;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Fibroblast gene expression following asthmatic bronchial epithelial cell conditioning correlates with epithelial donor lung function and exacerbation history
    Authors: SR Reeves, KA Barrow, TK Kolstad, MP White, LM Rich, TN Wight, JS Debley
    Sci Rep, 2018;8(1):15768.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. PFKFB4 Promotes Breast Cancer Metastasis via Induction of Hyaluronan Production in a p38-Dependent Manner
    Authors: R Gao, Y Liu, D Li, J Xun, W Zhou, P Wang, C Liu, X Li, W Shen, W Su, H Qiao, DG Stupack, N Luo
    Cell. Physiol. Biochem., 2018;50(6):2108-2123.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. PH20 is not expressed in murine CNS and oligodendrocyte precursor cells
    Authors: M Marella, J Ouyang, J Zombeck, C Zhao, L Huang, RJ Connor, KB Phan, MC Jorge, MA Printz, RD Paladini, AB Gelb, Z Huang, GI Frost, BJ Sugarman, L Steinman, G Wei, HM Shepard, DC Maneval, PJ Lapinskas
    Ann Clin Transl Neurol, 2017;4(3):191-211.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  20. Impaired Synthesis of Stromal Components in Response to Minnelide Improves Vascular Function, Drug Delivery, and Survival in Pancreatic Cancer.
    Authors: Banerjee S, Modi S, McGinn O, Zhao X, Dudeja V, Ramakrishnan S, Saluja A
    Clin Cancer Res, 2016;22(2):415-25.
    Species: Mouse
    Sample Types: Tissue Homogenates
  21. Transcription factor TBX4 regulates myofibroblast accumulation and lung fibrosis
    J Clin Invest, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Hyaluronan Does Not Regulate Human Epidermal Keratinocyte Proliferation and Differentiation
    Authors: J Malaisse, V Pendaries, F Hontoir, V De Glas, D Van Vlaend, M Simon, C Lambert de, Y Poumay, B Flamion
    J. Biol. Chem, 2016;291(12):6347-58.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. 4-methylumbelliferone diminishes catabolically-activated articular chondrocytes and cartilage explants via a mechanism independent of hyaluronan inhibition
    J Biol Chem, 2016;0(0):.
    Species: Human
    Sample Types: Cell Lysates
  24. Chronic lymphocytic leukemia: a paradigm of innate immune cross-tolerance.
    Authors: Jurado-Camino T, Cordoba R, Esteban-Burgos L, Hernandez-Jimenez E, Toledano V, Hernandez-Rivas J, Ruiz-Sainz E, Cobo T, Siliceo M, Perez de Diego R, Belda C, Cubillos-Zapata C, Lopez-Collazo E
    J Immunol, 2015;194(2):719-27.
    Species: Human
    Sample Types: Serum
  25. Articular Joint Lubricants during Osteoarthritis and Rheumatoid Arthritis Display Altered Levels and Molecular Species.
    Authors: Kosinska M, Ludwig T, Liebisch G, Zhang R, Siebert H, Wilhelm J, Kaesser U, Dettmeyer R, Klein H, Ishaque B, Rickert M, Schmitz G, Schmidt T, Steinmeyer J
    PLoS ONE, 2015;10(5):e0125192.
    Species: Human
    Sample Types: Synovial Fluid
  26. Inhibition of hyaluronan synthesis protects against central nervous system (CNS) autoimmunity and increases CXCL12 expression in the inflamed CNS.
    Authors: Mueller A, Yoon B, Sadiq S
    J Biol Chem, 2014;289(33):22888-99.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. Hyaluronan metabolism in human keratinocytes and atopic dermatitis skin is driven by a balance of hyaluronan synthases 1 and 3.
    Authors: Malaisse J, Bourguignon V, De Vuyst E, Lambert de Rouvroit C, Nikkels A, Flamion B, Poumay Y
    J Invest Dermatol, 2014;134(8):2174-82.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. Recombinant human hyaluronidase PH20 does not stimulate an acute inflammatory response and inhibits lipopolysaccharide-induced neutrophil recruitment in the air pouch model of inflammation.
    Authors: Huang Z, Zhao C, Chen Y, Cowell J, Wei G, Kultti A, Huang L, Thompson C, Rosengren S, Frost G, Shepard H
    J Immunol, 2014;192(11):5285-95.
    Species: Mouse
    Sample Types: Plasma
  29. Murine hyaluronidase 2 deficiency results in extracellular hyaluronan accumulation and severe cardiopulmonary dysfunction.
    Authors: Chowdhury, Biswajit, Hemming, Richard, Hombach-Klonisch, Sabine, Flamion, Bruno, Triggs-Raine, Barbara
    J Biol Chem, 2013;288(1):520-8.
    Species: Mouse
    Sample Types: Serum
  30. Molecular mass dependence of hyaluronan detection by sandwich ELISA-like assay and membrane blotting using biotinylated hyaluronan binding protein.
    Authors: Yuan, Han, Tank, Mihir, Alsofyani, Abeer, Shah, Naman, Talati, Nishant, LoBello, Jaclyn C, Kim, Jin Ryou, Oonuki, Yoji, de la Motte, Carol A, Cowman, Mary K
    Glycobiology, 2013;23(11):1270-80.
    Species: N/A
    Sample Types: Protein
  31. Effective targeting of the tumor microenvironment for cancer therapy.
    Authors: Jiang P, Li X, Thompson CB
    Anticancer Res., 2012;32(4):1203-12.
    Species: Human
    Sample Types: Cell Culture Supernates
  32. Deficiency of liver sinusoidal scavenger receptors stabilin-1 and -2 in mice causes glomerulofibrotic nephropathy via impaired hepatic clearance of noxious blood factors.
    Authors: Schledzewski K, Geraud C, Arnold B, Wang S, Grone HJ, Kempf T, Wollert KC, Straub BK, Schirmacher P, Demory A, Schonhaber H, Gratchev A, Dietz L, Thierse HJ, Kzhyshkowska J, Goerdt S
    J. Clin. Invest., 2011;121(2):703-14.
    Species: Mouse
    Sample Types: Plasma
  33. Peroxisome Proliferator-activated Receptor gamma Ligands Inhibit Transforming Growth Factor-beta-induced, Hyaluronan-dependent, T Cell Adhesion to Orbital Fibroblasts.
    Authors: Guo N, Woeller CF, Feldon SE, Phipps RP
    J. Biol. Chem., 2011;286(21):18856-67.
    Species: Human
    Sample Types: Cell Culture Supernates
  34. Chronic cigarette smoke exposure generates pathogenic T cells capable of driving COPD-like disease in Rag2-/- mice.
    Authors: Motz GT, Eppert BL, Wesselkamper SC, Flury JL, Borchers MT
    Am. J. Respir. Crit. Care Med., 2010;181(11):1223-33.
    Species: Mouse
    Sample Types: BALF
  35. Mast cell-derived prostaglandin D2 controls hyaluronan synthesis in human orbital fibroblasts via DP1 activation: implications for thyroid eye disease.
    Authors: Guo N, Baglole CJ, O'Loughlin CW, Feldon SE, Phipps RP
    J. Biol. Chem., 2010;285(21):15794-804.
    Species: Human
    Sample Types: Cell Culture Supernates
  36. Hyaluronan synthesis mediates the fibrotic response of keratocytes to transforming growth factor beta.
    Authors: Guo N, Li X, Mann MM, Funderburgh ML, Du Y, Funderburgh JL
    J. Biol. Chem., 2010;285(42):32012-9.
    Species: Bovine
    Sample Types: Cell Culture Supernates
  37. CD44 deficiency is associated with increased bacterial clearance but enhanced lung inflammation during Gram-negative pneumonia.
    Authors: van der Windt GJ, Florquin S, de Vos AF
    Am. J. Pathol., 2010;177(5):2483-94.
    Species: Mouse
    Sample Types: BALF
  38. Imatinib mesylate and AMN107 inhibit PDGF-signaling in orbital fibroblasts: a potential treatment for Graves' ophthalmopathy.
    Authors: van Steensel L, Paridaens D, Schrijver B, Dingjan GM, van Daele PL, van Hagen PM, van den Bosch WA, Drexhage HA, Hooijkaas H, Dik WA
    Invest. Ophthalmol. Vis. Sci., 2009;50(7):3091-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  39. Platelet-derived hyaluronidase 2 cleaves hyaluronan into fragments that trigger monocyte-mediated production of proinflammatory cytokines.
    Authors: de la Motte C, Nigro J, Vasanji A, Rho H, Kessler S, Bandyopadhyay S, Danese S, Fiocchi C, Stern R
    Am. J. Pathol., 2009;174(6):2254-64.
    Species: Human
    Sample Types: Cell Culture Supernates
  40. High levels of hyaluronan in idiopathic pulmonary arterial hypertension.
    Authors: Aytekin M, Comhair SA, de la Motte C, Bandyopadhyay SK, Farver CF, Hascall VC, Erzurum SC, Dweik RA
    Am. J. Physiol. Lung Cell Mol. Physiol., 2008;295(5):L789-99.
    Species: Human
    Sample Types: Plasma
  41. Interleukin (IL)-4, IL-13, and IL-17A differentially affect the profibrotic and proinflammatory functions of fibrocytes from asthmatic patients.
    Authors: Bellini A, Marini MA, Bianchetti L, Barczyk M, Schmidt M, Mattoli S
    Mucosal Immunol, 0;5(2):140-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  42. Biglycan, a danger signal that activates the NLRP3 inflammasome via toll-like and P2X receptors.
    Authors: Babelova A, Moreth K, Tsalastra-Greul W, Zeng-Brouwers J, Eickelberg O, Young MF, Bruckner P, Pfeilschifter J, Schaefer RM, Schaefer R, Grone HJ, Schaefer L
    J Biol Chem, 0;284(36):24035-48.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  43. 4-Methylumbelliferone Diminishes Catabolically Activated Articular Chondrocytes and Cartilage Explants via a Mechanism Independent of Hyaluronan Inhibition.
    Authors: Ishizuka S, Askew E, Ishizuka N, Knudson C, Knudson W
    J Biol Chem, 0;291(23):12087-104.
    Species: Human
    Sample Types: Cell Culture Supernates
  44. TGFbeta2-mediated production of hyaluronan is important for the induction of epicardial cell differentiation and invasion.
    Authors: Craig EA, Craig E, Austin A, Austin AF, Vaillancourt RR, Vaillancourt R, Barnett JV, Camenisch T, Camenisch TD
    Exp Cell Res, 0;316(20):3397-405.
    Species: Mouse
    Sample Types: Cell Culture Supernates

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Hyaluronan DuoSet ELISA
By Carmen Martin-Ruiz on 11/03/2021
Sample Tested: Cell culture supernatant

Hyaluronan DuoSet ELISA
By Christian Cassel on 12/15/2020
Sample Tested: Cell culture supernatant