Duoset kit

Manufactured by R&D Systems

Sourced in United States, United Kingdom, Germany, Italy

DuoSet kits are a series of ELISA development reagent kits designed for the quantitative measurement of specific proteins. The kits provide the components necessary to develop a sandwich ELISA, including capture and detection antibodies, and calibrated standards.

Automatically generated - may contain errors

Lab products found in correlation

Lipopolysaccharides (lps), by Merck Group (6 mentions) Ifn γ, by Merck Group (5 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (4 mentions) Protease inhibitor cocktail, by Roche (4 mentions) Elisa kit, by Thermo Fisher Scientific (3 mentions) Milliplex multiplex assay, by Merck Group (3 mentions) Opteia il 1β elisa set, by BD (3 mentions) 3 3 5 5 tetramethylbenzidine (tmb), by Merck Group (2 mentions) Sa hrp, by Corning (2 mentions) Mouse ccl3 mip 1α duoset elisa, by R&D Systems (2 mentions) Mouse ccl4 mip 1β duoset elisa, by R&D Systems (2 mentions) Fluorokine 1 multianalyte profiling kits, by R&D Systems (2 mentions) Cytometric bead array kit, by BD (2 mentions) Lysing matrix d, by MP Biomedicals (2 mentions) Quantikine kit, by R&D Systems (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Synergy ht, by Agilent Technologies (2 mentions) Quantikine, by R&D Systems (2 mentions) Opteiatm kits, by BD (2 mentions) Bradford assay, by Bio-Rad (2 mentions)

260 protocols using duoset kit

Biomarker Measurement from Serum Samples

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Serum aliquots from peripheral venipuncture samples were frozen and stored at −80C until shipment on dry ice to the UW Pulmonary Research Laboratory in Seattle, WA. Samples were tested for endothelial activation, inflammation, and immune activation biomarkers. Notably, the biomarkers we measured are stable at room temperature for ≥24 hours and resistant to up to four freeze-thaw cycles,^{29 (link)–31 (link)} and this testing constituted the first thaw for each aliquot used.
Serum concentrations of sTREM-1 and endothelin-1 were measured by ELISA (R&D Systems Duoset kits, Minneapolis, MN) according to manufacturer’s instructions using 50 μL of sample per well. Concentrations were interpolated from 4-parameter-fit standard curves. Serum concentrations of IL-6, CRP, SAA, sCD14, sCD163, sVCAM-1, sICAM-1, Ang-1, and Ang-2 were measured by Meso Scale Discovery assays (Meso Scale Diagnostics [MSD], Rockville, MD) according to manufacturer’s instructions. All assays except for sCD14 and sCD163 were standard MSD assays. The sCD14 and sCD163 assays were custom plates manufactured using a DuoSet kit from R&D. Samples were diluted to fit the dynamic ranges of the assays.

Attia E.F., Bhatraju P.K., Triplette M., Kosamo S., Maleche-Obimbo E., West T.E., Richardson B., Zifodya J.S., Eskander S., Njiru C.D., Warui D., Kicska G.A., Chung M.H., Crothers K., Liles W.C, & Graham S.M. (2020). Endothelial activation, innate immune activation, and inflammation are associated with post-bronchodilator airflow limitation and obstruction among adolescents living with HIV. Journal of acquired immune deficiency syndromes (1999), 83(3), 267-277.

+ Open protocol

+ Expand

Cytokine Quantification Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Culture supernatants were used to quantify cytokine expression according to manufacturers’ instructions for human TNF-α (Thermo Fisher Scientific, Affymetrix eBioscience, Waltham, MA, USA; Ready-Set-Go kit; sensitivity: 4–500 pg/mL), human CXCL10/IP-10 (R&D Systems, DuoSet kit; sensitivity: 31.2–2,000 pg/mL), and sIL-6Rα (R&D systems, DuoSet kit; sensitivity: 15.6–1,000 pg/mL). All supernatants were either undiluted or diluted 1:2 and 1:5 and were run on the same plate to ensure the readings were comparable. Absorbance was measured with the BioTek ELx800 Microplate Reader (Winooski, VT, USA) at 450 nm. Graphic data present the mean ± SEM, and n values for each experiment are indicated in figure legends.

Petes C., Mariani M.K., Yang Y., Grandvaux N, & Gee K. (2018). Interleukin (IL)-6 Inhibits IL-27- and IL-30-Mediated Inflammatory Responses in Human Monocytes. Frontiers in Immunology, 9, 256.

+ Open protocol

+ Expand

ATP-induced IL-1β release in J774A.1 cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

J774A.1 cells were seeded in 96-well plates at a concentration of 10⁵ cells/well and incubated overnight. Cells were primed for 4 h with 1 μg/ml E. coli O111:B4 LPS and subsequently stimulated for 30 min with 3 mM ATP in 100 μl medium. Where indicated, cells were treated with 100 μM BAPTA-AM (Tocris, Minneapolis, MN, USA) for 15 min before ATP treatment. Supernatants were collected and released IL-1β was evaluated with ELISA using the R&D Systems DuoSet kit according to the manufacturer's protocol. Developed plates were read on a Biotek Synergy H4 mutli-mode plate reader with Gen5 software.

Yaron J.R., Gangaraju S., Rao M.Y., Kong X., Zhang L., Su F., Tian Y., Glenn H.L, & Meldrum D.R. (2015). K+ regulates Ca2+ to drive inflammasome signaling: dynamic visualization of ion flux in live cells. Cell Death & Disease, 6(10), e1954-.

+ Open protocol

+ Expand

Macrophage Inflammation Modulation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

RAW 264.7 macrophage cells or BMDMs were plated at 2 × 10⁵ cells per well in 24-well plates and then incubated with or without LPS (1 μg/mL) in the absence or presence of various concentrations of samples. Dilutions of the cell culture medium were assayed for PGE₂, TNF-α, IL-6, and IL-1β. PGE₂ levels in cell culture medium were determined using a colorimetric competitive enzyme-linked immunosorbent assay (ELISA) kit (Enzo Life Science, NY, USA) according to manufacturer's instructions. TNF-α and IL-6 levels in cell culture medium were quantified using mouse DuoSet kit (R&D Systems, MN, USA) according to manufacturer's instructions.

Han H.S., Jang E., Shin J.S., Inn K.S., Lee J.H., Park G., Jang Y.P, & Lee K.T. (2017). Kyungheechunggan-Tang-01, a New Herbal Medication, Suppresses LPS-Induced Inflammatory Responses through JAK/STAT Signaling Pathway in RAW 264.7 Macrophages. Evidence-based Complementary and Alternative Medicine : eCAM, 2017, 7383104.

+ Open protocol

+ Expand

Quantifying Cytokine Levels in Pancreatic Cancer

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Equal numbers (1 × 10⁶) of cells CD18/HPAF-scram (control), CD18/HPAF-shKRAS, HPNE, HPNE-KRAS, E6-E7-st, E6-E7-st-KRAS, E6-E7-st-KRAS-NSC, E6-E7-st KRAS-shCXCR2 and KRAS-PDAC cells were plated in 60 mm dishes in complete medium. After attachment of cells to the plate, the medium was changed to serum-free DMEM. Supernatants of cultured cells were collected at 24 hours or 72 hours. Protein was isolated from tumors by homogenizing in a bullet blender using Mammalian Protein Extraction Reagent (Pierce, Rockford, IL) as a lysis buffer. ELISA assays for hCXCL8 and hCXCL1 were performed as described previously [39 (link)]. hCXCL5, mCXCL2, mCXCL5 and mCXCL7 ELISAs were performed using a duoset kit (R&D Systems, Minneapolis, MN) according to the manufacturer's protocol. All the experiments were performed in duplicates.

Purohit A., Varney M., Rachagani S., Ouellette M.M., Batra S.K, & Singh R.K. (2016). CXCR2 signaling regulates KRAS(G12D)-induced autocrine growth of pancreatic cancer. Oncotarget, 7(6), 7280-7296.

+ Open protocol

+ Expand

IL-10 Release from Degradable Hydrogel

Check if the same lab product or an alternative is used in the 5 most similar protocols

Composite hydrogels were formed with a high concentration of degradable microgels (50 μL bulk gel, n=3) and release buffer (1% bovine serum albumin in PBS) was added at 50 μL bulk gel/1 mL release buffer. Every other day, release buffer was collected via centrifugation at 1200 × g for 15 minutes, replaced, and stored at −20°C. After 2 weeks, composite hydrogels were fully degraded in hyaluronidase (1 mg/mL) and IL-10 release was quantified via ELISA (DuoSet kit, R&D Systems).

Chen M.H., Chung J.J., Mealy J.E., Zaman S., Li E.C., Arisi M.F., Atluri P, & Burdick J.A. (2018). Injectable supramolecular hydrogel/microgel composites for therapeutic delivery. Macromolecular bioscience, 19(1), e1800248.

+ Open protocol

+ Expand

Serum OPG Quantification by ELISA

Check if the same lab product or an alternative is used in the 5 most similar protocols

The serum OPG was determined in duplicate by ELISA with Duoset kit (DY805; R&D Systems, Minneapolis, MN) as recommended by the manufacturer. The ELISA system had an intraassay coefficient of variation of 3–9 % and an interassay coefficient of variation of 3–10 %, respectively.

Niu Y., Yang Z., Li X., Zhang W., Lu S., Zhang H., Chen X., Zhu L., Xing Y., Ning G., Qin L, & Su Q. (2015). Association of osteoprotegerin with impaired glucose regulation and microalbuminuria: the REACTION study. BMC Endocrine Disorders, 15, 75.

+ Open protocol

+ Expand

Growth Factor and Cytokine Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Levels of human basic fibroblast growth factor (bFGF), TGF-β1, interleukin (IL)-1β, tumor necrosis factor alpha (TNF-α), and pro-collagen I and mouse levels of TGF-β1, IL-1β, IL-6, and vascular endothelial growth factor (VEGF) were measured in the cell-free supernatants using a Duo-Set Kit (R&D Systems, Minneapolis, MI, USA), according to the manufacturer’s instructions. Results were expressed as pg/ml and determined using standard curves established for each assay.

Almeida Donanzam D.D., Donato T.A., dos Reis K.H., da Silva A.P., Finato A.C., dos Santos A.R., Cavalcante R.S., Mendes R.P, & Venturini J. (2020). Exoantigens of Paracoccidioides spp. Promote Proliferation and Modulation of Human and Mouse Pulmonary Fibroblasts. Frontiers in Cellular and Infection Microbiology, 10, 590025.

+ Open protocol

+ Expand

Cytokine Quantification by ELISA

Check if the same lab product or an alternative is used in the 5 most similar protocols

Interleukin (IL)-1β, IL-6, and TNF-α were measured by sandwich ELISA in the cell-culture supernatants using paired antibodies or Duoset kit (Biotechne R&D Systems, Minneapolis, MN, USA), IL-8/CXCL8 (Sigma–Biotechne R&D Systems, Minneapolis, MN, USA). IFN-α was also quantified by ELISA (PBL Assay Sciences, Piscataway, NJ, USA).

Petitjean S.J., Lecocq M., Lelong C., Denis R., Defrère S., Mariage P.A., Alsteens D, & Pilette C. (2022). Salvia miltiorrhiza Bunge as a Potential Natural Compound against COVID-19. Cells, 11(8), 1311.

+ Open protocol

+ Expand

Macrophage TNF-α Response to L. prolificans

Check if the same lab product or an alternative is used in the 5 most similar protocols

J774 macrophages were plated in 24-wells plates (2.0 × 10⁵ cells/ml/well) and after adhesion, stimulated in RPMI medium with addition of L. prolificans PRM (100 μg/ml) or LPS (O111:B4, 1 μg/ml). After stimulation for 1, 2 or 3 h, the supernatants were recovered for TNF-α determination by ELISA (R&D Systems’s DuoSet kit) according to the manufacturer’s instructions. In order to rule out the possibility that the stimulating activity was due to contaminating lipopolysaccharides, polymyxin B (10 μg/ml) was added 5 min prior to the addition of the stimulus. After incubation, supernatants were harvested, centrifuged at 12000 rpm for 10 min to remove cell debris and immediately measured by ELISA.

Xisto M.I., Santos S.S., Rossato L., Yoshikawa F.S., Haido R.M., de Almeida S.R, & Barreto-Bergter E. (2020). Peptidorhamnomannan from Lomentospora prolificans modulates the inflammatory response in macrophages infected with Candida albicans. BMC Microbiology, 20, 245.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!