Duoset elisa kit

Manufactured by R&D Systems

Sourced in United States, United Kingdom, Germany, France, Canada, Italy, Australia

DuoSet ELISA kits are laboratory reagent kits used for the quantitative measurement of specific proteins in a variety of sample types. The kits include a matched antibody pair and other necessary reagents to perform a sandwich enzyme-linked immunosorbent assay (ELISA). The assay measures the concentration of the target analyte in the sample.

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Close spelling variants of this product

DuoSet ELISA (635 citations) ELISAs (144 citations) DuoSets (49 citations) TNF-α DuoSet ELISA kits (7 citations) R&D Duoset ELISA kits (6 citations) Duoset cytokine ELISA kits (6 citations) Duoset ELISA kits for IL-1β (4 citations) Mouse IL-1β/IL-1F2 DuoSet ELISA kit (4 citations) DuoSets ELISA kits (3 citations) DuoSet enzyme linked immunosorbent assays (ELISA) kits (3 citations)

1 131 protocols using duoset elisa kit

Unveiling PSTI's Mechanisms: Apoptosis, Angiogenesis, and Barrier Function

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Having shown that PSTI reduced cell and organ damage in our in vitro and in vivo studies, samples from in vitro cell viability studies and whole animal I/R tissues were further analysed to examine PSTI’s possible modes of action.
Cell apoptosis assays
Active caspase-3 and active caspase-9 were determined using methods described previously [18 (link)], using commercial colorimetric assay kits (BF3100 and BF10100, R&D Systems). Concentrations of the anti-apoptotic protein Bcl2 and the pro-apoptotic protein Baxα were determined in the same cell lysates as used for caspase analyses, using Duoset Elisa kits (R&D Systems Europe Ltd).
Measurements of HIF1α, VEGF, Hsp70 and ICAM-1
HIF1α, VEGF, Hsp70 and ICAM1 concentration in the cleared cell lysates was determined using Duoset Elisa kits as per the manufacturer’s instructions (R&D Systems Europe). Hsp70 concentrations were determined using our previously published methods [18 (link)] with a Duoset Elisa kit (R&D Systems Europe).
Tight junction proteins
ZO1 and Claudin1 concentrations were determined using previously published methods [18 (link)] and standard ELISA kits (tight junction antibody samples pack 90–1200, Invitrogen)

Playford R.J, & Marchbank T. (2020). Pancreatic secretory trypsin inhibitor reduces multi-organ injury caused by gut ischemia/reperfusion in mice. PLoS ONE, 15(1), e0227059.

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Cytokine Secretion by NP-Exposed Cells

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The 48-well culture plates (Costar, France) were coated with a solution of 0.3% BSA (bovine serum albumin, Sigma-Aldrich, France) in DMEM/F12 (without phenol red, containing Hepes) and incubated for 15 min at 37 C and 5% CO 2 . After rinsing for three times with the culture media, 150 ml of cytokines at 500 pg/ ml (IL-6, IL-8, and GM-CSF standards from DuoSet Õ ELISA Kit, R&D Systems, Lille, France) were added in culture media. A total of 150 ml of NP dispersions was added to obtain a concentration of 75 mg/cm 2 (237.5 mg/ml) before incubation for 24 h at 37 C and 5% CO 2 . Supernatants were centrifuged at 10,000 rpm for 10 min at 4 C and supernatants stored at À80 C. Concentration of cytokines in the supernatants was evaluated using the DuoSet Õ ELISA Kit (R&D Systems). The OD was determined at 450 nm by a multiwell plate reader (Dynex, MRX; Thermolab systeme, Cergy Pontoise, France).

Guadagnini R., Halamoda Kenzaoui B., Walker L., Pojana G., Magdolenova Z., Bilanicova D., Saunders M., Juillerat-Jeanneret L., Marcomini A., Huk A., Dusinska M., Fjellsbø L.M., Marano F, & Boland S. (2015). Toxicity screenings of nanomaterials: challenges due to interference with assay processes and components of classic in vitro tests. Nanotoxicology, 9 Suppl 1.

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Melatonin and IL-1β Modulate Urokinase Pathway

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Briefly, 1 × 10⁵ cells were inoculated in 24-well culture wells in DMEM containing 10% FBS. After 24 h of attachment, the medium was changed to fresh medium with various concentrations of melatonin (dose study) or IL-1β. In some experiments, melatonin (50, 100, or 200 μg/mL, as indicated) or several signaling inhibitors (such as MEK/ ERK inhibitor [U0126], p38 MAPK inhibitor [SB203580], PI3K/Akt inhibitor [LY294002], and TAK1 inhibitor [5Z-7oxozeaenol]) were added before the addition of IL-1β (5 ng/mL), and cells were further incubated for 24 h. The culture medium was collected for enzyme-linked immunosorbent assay (ELISA) analysis of uPA (DuoSet ELISA kit, DY1310, R&D Systems Inc., Minneapolis, MN, USA), suPAR (DuoSet ELISA kit, DY807, R&D Systems), and PAI-1 (PAI-1 ELISA kit, 900-K383, PeproTech Company, Rocky Hill, NJ, USA) based on the manufacturer’s instructions.

Chang M.C., Zhong B.H., Lee H.N., Chuang F.H., Lee M.S., Chang H.H., Pan Y.H, & Jeng J.H. (2022). Melatonin exerts anti-fibrinolytic effects by regulating IL-1β-induced changes in uPA, uPAR, and PAI-1 expression/production in human dental pulp cells. Journal of Food and Drug Analysis, 30(3), 466-478.

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Quantification of immune proteins in nasal tissues

Cited 2 times

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Oncostatin M protein was detected in neutrophil cell culture supernatants using a DuoSet ELISA kit (R&D Systems, Minneapolis, MN). FSTL1 and GM-CSF protein were detected in neutrophil cell culture supernatants and FSTL1 was detected in sinus tissue extracts using DuoSet ELISA kits (R&D Systems). GM-CSF protein was detected in sinus tissue lysates using a Human Magnetic Protein Luminex Assay (R&D Systems). OSM protein was detected in sinus tissue lysates as previously described^{8 (link)}. Neutrophil Elastase protein (Hycult Biotech, Plymouth Meeting, PA) and Eosinophilic Cationic Protein (MBL International, Woburn, MA) were detected using ELISA, and sinus tissue extracts were prepared as previously described^{35 (link)}.

Pothoven K.L., Norton J.E., Suh L.A., Carter R.G., Harris K.E., Biyasheva A., Welch K., Shintani-Smith S., Conley D.B., Liu M.C., Kato A., Avila P.C., Hamid Q., Grammer LC I.I.I., Peters A.T., Kern R.C., Tan B.K, & Schleimer R.P. (2016). Neutrophils are a major source of the epithelial barrier disrupting cytokine Oncostatin M in mucosal airways disease. The Journal of allergy and clinical immunology, 139(6), 1966-1978.e9.

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VEGF and FGF-2 Quantification

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Secreted levels of Human VEGF were determined by DuoSet ELISA Kit (R&D System, USA) from free-cell culture supernatants collected after treatments. FGF-2 expression levels were determined by DuoSet ELISA Kit (R&D System, USA) from protein extracts. The assays were carried out according to instructions provided by the manufacturer.

Vitale D.L., Caon I., Parnigoni A., Sevic I., Spinelli F.M., Icardi A., Passi A., Vigetti D., & Alaniz L. (2020). Knockdown Of Udp-Glucose Dehydrogenase Facilitates Epirubicin-Resistance In MDA-MB-231 Breast Cancer Cells By Regulation Of Hyaluronan Synthesis.

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Interferon Response in MEFs and HEK293Ts

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MEFs were seeded in DMEM with 2% FBS and HEK293Ts were seeded in DMEM with 10% FBS. After 18 h, cells were mock-transfected or transfected with 5 µg/mL HMW PolyIC (Invivogen) using Lipofectamine 2000 (Thermofisher) for 7 h or infected with 40 HAU/mL SeV for 4.5 or 24 h where stated. The culture medium was cleared by centrifugation at 17,000× g and stored at −20 °C before analysis by ELISA. The level of human or mouse CXCL10/IP-10 was determined using a DuoSet ELISA kit (R&D Systems, Minneapolis, MN, USA) and the level of mouse IL-6 was determined using a DuoSet ELISA kit (R&D systems) following the manufacturer’s instructions. Data were collected and analysed using the MARS data analysis software on the FLUOstar Omega Luminometer (BMG Labtech, Aylesbury, UK). Experiments were carried out in triplicate and measured with technical repeats, unless stated otherwise. RNA extraction, cDNA synthesis, and RT-qPCR were carried out as described previously using first strand synthesis (Invitrogen) [35 (link)]. qPCR was performed using the primers indicated in Table S2.

Pallett M.A., Lu Y, & Smith G.L. (2022). DDX50 Is a Viral Restriction Factor That Enhances IRF3 Activation. Viruses, 14(2), 316.

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IL-37b Modulates LPS-Induced Inflammation

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RAW cells (ATCC TIB-71^TM) were seeded into 96-well plates with 5 × 10⁴/well in DMEM with 10% FBS and cultured overnight. Human PBMC were purified from whole blood samples by Ficoll-Paque PLUS (GE Healthcare. Illinois, USA) and seeded into 96-well plates with 1 × 10⁵ in RMPI RPMI 1460 containing 10% FBS, 2 mM glutamine, 1% nonessential amino acids, 1% sodium pyruvate, and 50 U/ml penicillin and streptomycin. After overnight culture, these cells were pretreated with gradient concentrations of recombinant IL-37b from 300 ng/ml to 0.3 ng/ml for 24 h. Then, the cells were changed into fresh culture medium with same concentration of recombinant IL-37b following adding 50 ng/ml E.coli LPS. After 8 h LPS stimulation, culture supernatants were collected and level of IL-6 in the supernatant of RAW cells were determined by DuoSet ELISA kit (DY406, R&D System) and level of IL-6 in the supernatant of human PBMC were determined by DuoSet ELISA kit (DY206, R&D System).

Offenbacher S., Jiao Y., Kim S.J., Marchesan J., Moss K.L., Jing L., Divaris K., Bencharit S., Agler C.S., Morelli T., Zhang S., Sun L., Seaman W.T., Cowley D., Barros S.P., Beck J.D., Munz M., Schaefer A.S, & North K.E. (2018). GWAS for Interleukin-1β levels in gingival crevicular fluid identifies IL37 variants in periodontal inflammation. Nature Communications, 9, 3686.

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Quantification of Inflammatory Markers

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The concentration of IL-6 in the supernatant of THP-1 cells was determined using the DuoSet ELISA kit (R&D, DY206) according to the manufacturer’s instructions. The concentration of CRP, IL-6, and IL-1β levels in the serum of WT and Gaplinc-KO mice was determined using the DuoSet ELISA kits (R&D, DY1829, DY406, and DY401) according to the manufacturer’s instructions. The concentration of lactate levels in the serum of WT and Gaplinc-KO mice was determined using l-Lactate Assay Kit (Abcam, ab65331).

Vollmers A.C., Covarrubias S., Kuang D., Shulkin A., Iwuagwu J., Katzman S., Song R., Viswanathan K., Vollmers C., Wakeland E, & Carpenter S. (2021). A conserved long noncoding RNA, GAPLINC, modulates the immune response during endotoxic shock. Proceedings of the National Academy of Sciences of the United States of America, 118(7), e2016648118.

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Quantifying Cytokine and CRP Levels

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The concentrations of IL-1β, IL-6, TNFα, IL-10, IL-17A and IL-22 were measured using MSD V-Plex Proinflammatory Panel 1 and Th17 Panel 1 kits (Meso Scale Diagnostics, Rockville, MD). The concentration of total TGFβ1 was measured using a Duo Set ELISA kit following acid activation and neutralization (R&D Systems, Minneapolis, MN). The concentration of C-Reactive Protein (CRP) was measured using a Duo Set ELISA kit from R&D Systems, Minneapolis, MN.

Fernandez-Botran R., Plankey M.W., Ware D, & Bordon J. (2021). Changes in liver steatosis in HIV-positive women are associated with the BMI, but not with biomarkers. Cytokine, 144, 155573.

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Serum and Supernatant ELISA Quantification

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Human TNFα was quantified in blood serum (1:10 dilution) using a DuoSet ELISA kit (R&D Systems, Minneapolis, MN). Human IFNγ and IL10 were quantified in culture supernatants (1:2 dilution) using DuoSet ELISA kits (R&D Systems). DuoSet ELISA Ancillary Reagent Kit 2 was used in conjunction with all DuoSet ELISA kits. Human IgG and IgM was quantified in blood serum (1:1000 dilution) and culture supernatants (1:10 dilution) using the Human IgG/IgM total ELISA Ready-SET-Go! Kits (Invitrogen/eBioscience). All ELISAs were performed as per manufacturer’s protocols. Plates were read at 450 nm wavelength on a SpectraMAX190 and analyzed using the SoftMax Pro software (Molecular Devices, San Jose, CA).

Carpenter R.S., Jiang R.R., Brennan F.H., Hall J.C., Gottipati M.K., Niewiesk S, & Popovich P.G. (2019). Human immune cells infiltrate the spinal cord and impair recovery after spinal cord injury in humanized mice. Scientific Reports, 9, 19105.

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