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Hyaluronan duoset elisa

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The Hyaluronan DuoSet ELISA is a laboratory equipment product that enables the quantitative measurement of hyaluronan levels in various sample types. It is a sandwich enzyme-linked immunosorbent assay (ELISA) designed to detect and quantify hyaluronan, also known as hyaluronic acid, in biological samples.

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Lab products found in correlation

Stains all, by Merck Group (2 mentions) Simoa assay, by Quanterix (1 mentions) Human fabp2 1 fabp duoset elisa, by R&D Systems (1 mentions) Human cxcl10 ip 10 duoset elisa, by R&D Systems (1 mentions) Human cd163 duoset elisa, by R&D Systems (1 mentions) Human cd14 duoset elisa, by R&D Systems (1 mentions) Proteinase k, by Roche (1 mentions) Versadoc imaging system, by Bio-Rad (1 mentions) L lactate assay kit 1, by Eton Bioscience (1 mentions) Proteinase k recombinant pcr grade, by Roche (1 mentions) Cytation 1, by Agilent Technologies (1 mentions)

Close spelling variants of this product

DuoSet ELISA (635 citations) ELISAs (144 citations) DuoSets (49 citations) Hyaluronan (14 citations) Hyaluronan DuoSet ELISA kit (13 citations) Hyaluronan DuoSet (11 citations)

9 protocols using hyaluronan duoset elisa

1

Biomarkers of Immune Activation in HIV

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We centrally measured plasma levels of ten soluble biomarkers of all participants. sTNFRII, sCD14, sCD163, CXCL10, I-FABP, and hyaluronic acid were measured by specific ELISA (Human TNF RII/TNFRSF1B DuoSet ELISA, Human CD14 DuoSet ELISA, Human CD163 DuoSet ELISA, Human CXCL10/IP10 DuoSet ELISA, Human FABP2/I-FABP DuoSet ELISA, and Hyaluronan DuoSet ELISA, R&D Systems). IL-17, IL-6, and TNF-α were measured by single-molecule array (SiMoA) assay (Quanterix), and CRP by immunochemistry (CRP LX HS, Cobas C, integra, Roche Diagnostics). Samples with undetectable levels were attributed half the threshold value.
We measured plasma HIV RNA levels of PRIMO participants, using an ultrasensitive real-time PCR technique (GENERIC HIV, Biocentric, France) and total HIV DNA levels from whole blood samples using a real-time PCR assay (GENERIC Biocentric, France) [19] (link).
The data that support the findings of this study are available on request to the corresponding author. The data are not publicly available due to privacy restrictions.
Novelli S., Lécuroux C., Goujard C., Reynes J., Villemant A., Blum L., Essat A., Avettand-Fenoël V., Launay O., Molina J.M., Bourgeois C, & Meyer L. (2020). Persistence of monocyte activation under treatment in people followed since acute HIV-1 infection relative to participants at high or low risk of HIV infection. EBioMedicine, 62, 103129.
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2

Quantification of Hyaluronan in Equine Synovial Fluid

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A commercially available sandwich ELISA (Hyaluronan DuoSet ELISA,
R&D Systems, Minneapolis, USA) was used to quantify HA concentration in
synovial fluid. This assay utilizes recombinant human aggrecan as the HA
capture reagent and biotinylated recombinant human aggrecan to detect bound
HA using streptavidin conjugated to horseradish peroxidase. Synovial fluid
samples from all horses at all time points were diluted 1:80,000 in 5% Tween
20 in PBS and measured in duplicate. Absorbance was measured at 450nm with
wavelength correction set at 540nm. HA MW distribution was determined by gel
electrophoresis as previously described29 (link). Briefly, synovial fluid samples were diluted (1:20
in PBS) and treated overnight with proteinase K (Roche Applied Science,
Mannheim, Germany) prior to electrophoresis (50V for 8 hours) on a 1%
agarose gel. HiLadder (0.5–1.5 MDa) and Mega-HA Ladder
(1.5–6.1 MDa) (Amsbio LLC, Cambridge, MA) were used for MW reference.
Gels were stained with 0.005% Stains-All (Sigma-Aldrich, St. Louis, MO) in
50% ethanol, then de-stained with a 10% ethanol solution prior to image
acquisition (Bio-Rad VersaDoc Imaging System, Hercules, CA). Where
necessary, gel images were combined using the Stitching plugin30 (link) prior to quantification of
band intensity with ImageJ software31 (link).
Peal B.T., Gagliardi R., Su J., Fortier L.A., Delco M.L., Nixon A.J, & Reesink H.L. (2020). Synovial Fluid Lubricin and Hyaluronan are Altered in Equine Osteochondral Fragmentation, Cartilage Impact Injury and Full-Thickness Cartilage Defect Models. Journal of orthopaedic research : official publication of the Orthopaedic Research Society, 38(8), 1826-1835.
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3

Evaluating Hyaluronan Production and Lactate Levels in IL-1β-Treated Cells

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Cells were incubated for 12 h with 0.1 ng/ml IL-1β in fresh serum-free culture medium with or without 1.0 mM 4-MU. The culture supernatant fluids were collected and the HA was quantified by enzyme-linked sandwich assay (Hyaluronan DuoSet ELISA, DY3614-05, R&D Systems) following the manufacturer’s instructions. The lactate concentration in culture supernatants was determined using l-Lactate Assay Kit I (Eton Bioscience, Research Triangle, NC, USA) according to the manufacturer’s instructions.
Idota M., Ishizuka S., Hiraiwa H., Yamashita S., Oba H., Kawamura Y., Sakaguchi T., Haga T., Mizuno T., Kawashima I., Kuriyama K, & Imagama S. (2021). 4-Methylumbelliferone suppresses catabolic activation in anterior cruciate ligament-derived cells via a mechanism independent of hyaluronan inhibition. Journal of Orthopaedic Surgery and Research, 16, 507.
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4

Quantifying Heparanase and Hyaluronan Levels

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ELISAs specific for Heparanase-1 (amsbio, #EA1340Hu) and Heparanase-2 (amsbio, #E5389Hu) were performed in plasma samples. All samples were measured using a Tecan absorbance microplate reader (Sunrise, #F039300) at 450 nm. All measurements were performed in duplicates. Hyaluronan concentration was measured using the Hyaluronan DuoSet ELISA according to manufacturer’s instruction (R&D Systems, Wiesbaden, Germany).
Stahl K., Hillebrand U.C., Kiyan Y., Seeliger B., Schmidt J.J., Schenk H., Pape T., Schmidt B.M., Welte T., Hoeper M.M., Sauer A., Wygrecka M., Bode C., Wedemeyer H., Haller H, & David S. (2021). Effects of therapeutic plasma exchange on the endothelial glycocalyx in septic shock. Intensive Care Medicine Experimental, 9, 57.
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5

Quantifying Aggrecan and Hyaluronan

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Aggrecan was measured in cell culture supernatant using the Aggrecan Human ELISA Kit (Thermo Fisher, Cat# KAP1461) according to the manufacturer’s protocol. HA was also measured in cell culture supernatant using the Hyaluronan DuoSet ELISA (R&D Systems, Cat# DY361405) according to the manufacturer’s protocol.
Krawetz R.J., Wu Y.E., Bertram K.L., Shonak A., Masson A.O., Ren G., Leonard C., Kapoor M., Matyas J.R, & Salo P.T. (2022). Synovial mesenchymal progenitor derived aggrecan regulates cartilage homeostasis and endogenous repair capacity. Cell Death & Disease, 13(5), 470.
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6

Synovial Fluid Hyaluronan Analysis

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Synovial fluid HA concentration was measured at all time points using a commercially available HA ELISA (Hyaluronan DuoSet ELISA, Cat#: DY3614–05, R&D Systems, Minneapolis, MN) [72 (link)]. The distribution of HA molecular weights was determined by gel electrophoresis in a similar manner to that described previously [73 (link)]. Synovial fluid samples were diluted 1:15 with phosphate buffered saline and incubated overnight with 75 μg/ mL proteinase k (Proteinase K, recombinant, PCR grade, Roche Applied Science, Mannheim, Germany). Samples and standards, HiLadder (0.5–1.5 MDa) and Mega-HA Ladder (1.5–6.1 MDa; AMS Biotechnology Limited, Cambridge, MA) were loaded onto a 0.5% agarose gel and run at 57 V for 8 h. Gels were stained for 24 h in 0.005% Stains-All (Sigma-Aldrich, St. Louis, MO) in 50% ethanol and de-stained in 10% ethanol for 24 h with final destaining occurring on exposure to ambient light. Images of gels acquired using a Bio-Rad VersaDoc Imaging System (Hercules, CA) with relative band intensity calculated using Fiji Software (ImageJ). The intra-assay coefficient of variation for the HA ELISA was 9.1%.
Watkins A., Fasanello D., Stefanovski D., Schurer S., Caracappa K., D’Agostino A., Costello E., Freer H., Rollins A., Read C., Su J., Colville M., Paszek M., Wagner B, & Reesink H. (2021). 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. BMC Veterinary Research, 17, 189.
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7

Quantification of Hyaluronic Acid Content

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Hyaluronic acid contents were measured using Hyaluronan DuoSet ELISA (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s protocol. Briefly, chopped dorsal skin tissue specimens or harvested HDF cells were lysed with RIPA buffer and quantified using the BCA assay. Equal amounts of protein and biotin-labeled antibody were simultaneously added to each well conjugated with the capture antibody against hyaluronic acid and reacted at 37 °C for 45 min. After washing the plate three times with 1× wash solution, HRP-Streptavidin conjugated working solution was added to each well and incubated at 37 °C for 30 min. After washing the plate five times with the wash solution, tetramethylbenzidine substrate was added to each well and reacted at 37 °C in the dark for 15 min. After stopping the reaction by adding stop solution to each well, the absorbance was measured at 450 nm using a Cytation 1 (BioTek, Winooski, VT, USA) at Biopolymer Research Center for Advanced Materials.
Sim W.J., Kim J., Baek K.S., Lim W, & Lim T.G. (2023). Porcine Placenta Peptide Inhibits UVB-Induced Skin Wrinkle Formation and Dehydration: Insights into MAPK Signaling Pathways from In Vitro and In Vivo Studies. International Journal of Molecular Sciences, 25(1), 83.
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8

Hyaluronic Acid Quantification Assay

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1x105 cells were seeded in 48-well plates and incubated for 24 h under appropriate treatments (argatroban addition or sh.E2F1, sh.MTA1 or sh.ctrl). Culture supernatants were collected and quantification of hyaluronic acid was performed by the enzyme- linked sandwich assay Hyaluronan DuoSet ELISA (R&D Systems, Minneapolis, MN, USA) following the manufacturer's instructions.
Goody D., Gupta S.K., Engelmann D., Spitschak A., Marquardt S., Mikkat S., Meier C., Hauser C., Gundlach J.P., Egberts J.H., Martin H., Schumacher T., Trauzold A., Wolkenhauer O., Logotheti S, & Pützer B.M. (2019). Drug Repositioning Inferred from E2F1-Coregulator Interactions Studies for the Prevention and Treatment of Metastatic Cancers. Theranostics, 9(5), 1490-1509.
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9

Quantification of Plasma Hyaluronan Levels

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Fasted venous blood samples were collected in ethylene diamine tetra acetic acid (EDTA) containing tubes and stored at − 80 °C until further analysis, as previously described41 (link). Plasma samples obtained at baseline, available for secondary research were used in the present study; only blood samples of subjects who provided written approval for use of body material for secondary research purposes were used. Natural plasma HA was measured using a solid phase HA binding protein-based sandwich enzyme-linked immunosorbent assay (ELISA), according to the manufacturer’s protocol (Hyaluronan DuoSet ELISA, R&D Systems, Minneapolis, MN, USA). Samples were measured in duplicate, and phosphate buffered saline samples were included and confirmed as negative controls. Results were analyzed in Excel (Microsoft Excel 2007, Redmond, WA, USA). The average intra-assay coefficient of variation was 12.6%. A power calculation is provided in the online supplement.
Waeijen-Smit K., Reynaert N.L., Beijers R.J., Houben-Wilke S., Simons S.O., Spruit M.A, & Franssen F.M. (2021). Alterations in plasma hyaluronic acid in patients with clinically stable COPD versus (non)smoking controls. Scientific Reports, 11, 15883.
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