Duoset enzyme linked immunosorbent assay elisa

Manufactured by R&D Systems

Sourced in United States

The DuoSet enzyme-linked immunosorbent assay (ELISA) is a laboratory equipment product offered by R&D Systems. It is designed to facilitate the quantitative measurement of specific proteins or other analytes in biological samples.

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Lab products found in correlation

6490 triple quadrupole mass spectrometer, by Agilent Technologies (1 mentions) Milliplex map high sensitivity human cytokine magnetic bead panel, by Merck Group (1 mentions) Unicel dxc 800, by Beckman Coulter (1 mentions) Hematology analyzer, by Sysmex (1 mentions) Streptavidin horseradish peroxidase, by R&D Systems (1 mentions) Softmax pro version 5, by Molecular Devices (1 mentions) Versamax, by Molecular Devices (1 mentions) Human primary osteoblasts, by Lonza (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)

6 protocols using duoset enzyme linked immunosorbent assay elisa

Comprehensive Plasma Lipid and Cytokine Analysis

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A panel of 51 eicosanoids and lipid metabolites were measured in plasma samples using a 6490 Triple Quadrupole mass spectrometer (Agilent, New Castle, DE, USA). The individual eicosanoids were identified using metabolite-specific fragmentation and retention times¹². We used the Milliplex MAP High Sensitivity Human Cytokine Magnetic Bead Panel (EMD Millipore Corp., St. Charles, MO) to measure four cytokines: IL-1β, IL-6, TNF-α, and IL-10. An additional inflammation marker, CRP, was also measured using a DuoSet enzyme-linked immunosorbent assay (ELISA) (R&D Systems, Minneapolis, MN)^{10 (link)}.
We measured three protein oxidation markers in plasma samples: 3-nitrotyrosine [NY], 3-chlorotyrosine [CY], and o,o’-dityrosine [DY]. To quantify these biomarkers, total plasma protein was first precipitated and diluted with a phosphate buffer. The samples were then delipidated, injected with isotopically labeled standards, and hydrolyzed for 24 h. Subsequently, the processed plasma samples underwent liquid chromatography-electrospray ionization tandem mass spectrometry. Additional oxidative stress markers 8-IP and 8-OHdG were measured in urine samples at Cayman Chemical (Ann Arbor, MI). 8-IP was quantified using affinity column chromatography followed by an enzyme immunoassay, and 8-OHdG was measured directly through an enzyme immunoassay⁴⁸.

Aung M.T., Song Y., Ferguson K.K., Cantonwine D.E., Zeng L., McElrath T.F., Pennathur S., Meeker J.D, & Mukherjee B. (2020). Application of an analytical framework for multivariate mediation analysis of environmental data. Nature Communications, 11, 5624.

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Evaluation of Iron Biomarkers

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All participants underwent blood collection by venipuncture at a medical facility. Hemoglobin (Hb) was measured by a complete blood cell count using a hematology analyzer (Sysmex, Kobe, Japan). Serum ferritin was measured by a chemiluminescence immunoassay by Beckman Coulter Unicel DxC 800 (Beckman Coulter, Brea, CA, USA). Transferrin saturation (TS) was calculated by dividing serum iron by the total iron-binding capacity (TIBC). Serum iron was measured by a ferrozine-based colorimetric assay and TIBC by an immunoturbidimetric method with a Beckman Coulter Unicel DxC 800 (Beckman Coulter). Serum hepcidin levels were measured with a human hepcidin DuoSet enzyme-linked immunosorbent assay (ELISA) (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s instructions. Variation in the intra- and inter-assay coefficients of serum hepcidin was <10%.

Mayasari N.R., Bai C.H., Hu T.Y., Chao J.C., Chen Y.C., Huang Y.L., Wang F.F., Tinkov A.A., Skalny A.V, & Chang J.S. (2021). Associations of Food and Nutrient Intake with Serum Hepcidin and the Risk of Gestational Iron-Deficiency Anemia among Pregnant Women: A Population-Based Study. Nutrients, 13(10), 3501.

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CRP and PTX3 Measurement in Plasma

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CRP was measured with high‐sensitivity technique (detection limit 0.15 mg/L), performed by turbidimetry, at the Clinical Chemistry laboratory of the University Hospital in Linköping.
A DuoSet enzyme-linked immunosorbent assay (ELISA; detection limit 0.22 µg/L) kit was used to analyze PTX3 levels in plasma (R&D Systems, Minneapolis, MN, USA). Assays were performed according to the manufacturers’ instructions as previously described (29 (link), 30 (link)). Briefly, Costar (Corning, NY, USA) half‐area plates were coated with 2 µg/mL of mouse anti‐human PTX3 and incubated overnight. Plates were blocked by 1% bovine serum albumin in PBS for 1 h and incubated thereafter with samples and standards for 2 h. Biotinylated goat anti‐human PTX3 (60 ng/mL) was added and incubated for 2 h, followed by addition of streptavidin horseradish peroxidase (R&D Systems) diluted 1:40 and another 20 min of incubation. Plates were developed with tetramethylbenzidine substrate and the reaction was stopped by adding 1 M H₂SO₄. All incubations were performed at room temperature. Plate reader VersaMax (Molecular devices, San Jose, CA, USA) and software SoftMax Pro version 5.4.1 (Molecular devices) were used.

Wirestam L., Pihl S., Saleh M., Wetterö J, & Sjöwall C. (2021). Plasma C-Reactive Protein and Pentraxin-3 Reference Intervals During Normal Pregnancy. Frontiers in Immunology, 12, 722118.

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Quantifying Cytokine and Chemokine Levels

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Human IL-1 beta/IL-1F2 DuoSet Enzyme-Linked Immunosorbent Assay (ELISA) (R&D #DY201) and Human CXCL10/IP-10 DuoSet ELISA (R&D #DY266) were performed according to the manufacturer’s recommendations.

Zhou J.Y., Sarkar M.K., Okamura K., Harris J.E., Gudjonsson J.E, & Fitzgerald K.A. (2023). Activation of the NLRP1 inflammasome in human keratinocytes by the dsDNA mimetic poly(dA:dT). Proceedings of the National Academy of Sciences of the United States of America, 120(5), e2213777120.

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Cytokine Quantification via ELISA

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TGF-β and TNF-α were measured in collected supernatants by Duoset enzyme-linked immunosorbent assay (ELISA) (R&D Systems Inc., Minneapolis, MN) according to the manufacturer protocol. Absorbencies were measured at 450 nm using the UV-max ELISA plate reader (SunriseTM). The ELISA reader-controlling software (Magellan, V 7.1) readily processes the digital data of raw absorbance value into a standard curve from which cytokine concentrations of unknown samples can be derived directly. Results are expressed as a pictogram of cytokine per milliliter of the supernatant.

El-Desoky A.M., Ali Y.B.M, & Talaat R.M. (2022). Cytotoxic effect of combining two antisense oligonucleotides against telomerase rna component (hTR and mRNA of centromere protein B (CENP-B) in hepatocellular carcinoma cells. Anais da Academia Brasileira de Ciencias, 94(2).

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Characterizing Lung Cancer-Osteoblast Interactions

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Chemicals. tcn was obtained from extrasynthese (genay, france), dissolved in dimethyl sulfoxide (DMSo) (Sigmaaldrich, St. louis, Mo, uSa), and stored at -20˚C. Control cultures received the carrier solvent (0.1% DMSo). all chemicals used were in their purest form available commercially.
Cell culture and conditioned medium. Human non-small cell lung cancer H460 cells were obtained from the american type culture collection (HtB-177) (Manassas, Va, uSa) and cultured in rPMi-1640 medium containing 10% fetal bovine serum (fBS) (both from gibco-Brl, gaithersburg, MD, uSa). Human primary osteoblasts were obtained from lonza (Walkersville, MD, uSa) and cultured in osteoblast medium (oBM) (lonza).
to obtain the various conditioned media (cM), H460 cells (2x10 6 /100 mm dish) were treated with various concentrations of BaP (Sigma-aldrich) for 6 h. after treatment, the medium was replaced and the supernatant harvested and filtered (0.22 mm) after 24 h of incubation.
Measurement of secreted factors. Supernatants from osteoblasts and H460 cells were collected. levels of osteoprotegerin (oPg), macrophage colony-stimulating factor (M-cSf), ranKl and il-8 were assessed and quantified using the DuoSet enzyme-linked immunosorbent assay (eliSa) (r&D Systems, Minneapolis, Mn, uSa). PtHrP levels were determined by an eliSa kit (abnova corp., taipei, taiwan).

Hung J.Y., Chang W.A., Tsai Y.M., Hsu Y.L., Chiang H.H., Chou S.H., Huang M.S, & Kuo P.L. (2015). Tricetin, a dietary flavonoid, suppresses benzo(a)pyrene‑induced human non‑small cell lung cancer bone metastasis. International journal of oncology, 46(5).

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