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Elisa ready set go

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ELISA Ready-SET-Go is a pre-coated enzyme-linked immunosorbent assay (ELISA) kit. It is designed to detect and quantify specific target analytes in biological samples.

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Lab products found in correlation

Duoset elisa, by R&D Systems (7 mentions) Facsdiva software, by BD (5 mentions) Platinum elisa, by Thermo Fisher Scientific (3 mentions) Duoset elisa kit, by R&D Systems (3 mentions) Griess reagent system, by Promega (2 mentions) Fv efluor 506, by Thermo Fisher Scientific (2 mentions) Annexin 5 fitc kit, by Thermo Fisher Scientific (2 mentions) Lsr 2 sorp flow cytometer, by BD (2 mentions) Th1 th2 th17 assay, by BD (2 mentions) Bio plex 200 system, by Bio-Rad (2 mentions) Elisa kit, by Thermo Fisher Scientific (2 mentions) Costar eia ria plates, by Corning (2 mentions) Opteia set, by BD (2 mentions) Tmb substrate solution, by BioLegend (2 mentions) Powerwavex, by Agilent Technologies (2 mentions) Multiskan go, by Thermo Fisher Scientific (2 mentions) Opteia elisa set, by BD (2 mentions) Golgistop, by BD (2 mentions) Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions) Legendplex, by BioLegend (1 mentions)

110 protocols using elisa ready set go

1

TLR Agonist-Induced Cytokine Profiling in Dendritic Cells

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CD4− MutuDC2s and MutuDC1s were seeded at a density of 2.5 × 105 cells/cm2 in 96-well or 48-well plates and incubated for 24 h with 450 μL/cm2 of the following TLR agonists diluted in complete medium: Pam3CSK4 (150 ng/mL, tlrl-pms, InvivoGen), poly(I:C) (8.5 μg/mL, tlrl-pic, InvivoGen), LPS from E. coli (100 ng/mL, tlrl-peklps, InvivoGen), ultrapure flagellin from B. subtilis (100 ng/mL, tlrl-pbsfla, InvivoGen), FSL-1 (100 ng/mL, tlrl-fsl, InvivoGen), Gardiquimod™ (1 μg/mL, tlrl-gdqs, InvivoGen), CpG ODN 1826 (1 μM, TriLink BIOTECHNOLOGIES). In all the experiments each condition was plated in technical triplicate. The supernatants were analyzed by ELISA for the presence of IL-6, IL-10, IL-12/IL-23 p40, IL-12p70, and MCP-1(CCL2) using the following kits according to manufacturer's instructions: Mouse IL-6 ELISA Set (555240, BD Biosciences) or Mouse IL-6 ELISA Ready-SET-Go! (88-7064, eBioscience), Mouse IL-10 ELISA Set (555252, BD Biosciences) or Mouse IL-10 (Interleukin-10) ELISA Ready-SET-Go! (88-7104, eBioscience), Mouse IL-12 (p40) ELISA Set (555165, BD Biosciences), Mouse IL-12 (p70) ELISA Set (555256, BD Biosciences), Mouse CCL2 (MCP-1) ELISA Ready-SET-Go! (88-7391, eBioscience).
Pigni M., Ashok D., Stevanin M, & Acha-Orbea H. (2018). Establishment and Characterization of a Functionally Competent Type 2 Conventional Dendritic Cell Line. Frontiers in Immunology, 9, 1912.
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2

IFN-γ Production in SVF Cells

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WT and p40−/− MAT and SAT SVF cells were
added to 96 well plates
(3,5 × 105 SVF
cells/well) and cultured for 48 h in complete RMPI alone or with
1,75 × 106 freeze-killed
NcT at a ratio cell:NcT of 1:5, prepared as described above. IFN-γ
levels in culture supernatants were quantified with Ready-Set-Go!®
ELISA (eBioscience) according to manufacturer’s instructions.
Teixeira L., Marques R.M., Ferreirinha P., Bezerra F., Melo J., Moreira J., Pinto A., Correia A., Ferreira P.G, & Vilanova M. (2016). Enrichment of IFN-γ producing cells in different murine adipose tissue depots upon infection with an apicomplexan parasite. Scientific Reports, 6, 23475.
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3

Protein Concentration Measurement Protocols

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Measurement of protein concentration was determined either by a standard ELISA or bead-based immunoassay (LEGENDplex; BioLegend) as indicated. ELISAs were performed according to the manufacturer's protocol (Ready-SET-Go ELISA; eBioscience) on supernatants from in vitro stimulations. Absorbance values were obtained at 450 nm using an Emax microplate reader (Molecular Devices), and IL-2 concentration was determined by standard curve. Protein concentration values were normalized to the number of sorted cells plated. LEGENDplex assays were performed according to the manufacturer’s protocols.
Williams J.B., Horton B.L., Zheng Y., Duan Y., Powell J.D, & Gajewski T.F. (2017). The EGR2 targets LAG-3 and 4-1BB describe and regulate dysfunctional antigen-specific CD8+ T cells in the tumor microenvironment. The Journal of Experimental Medicine, 214(2), 381-400.
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4

Cytokine Analysis by ELISA

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The cytokines in culture supernatants were assessed using "sandwich" ELISA (Ready-Set-Go ELISA and Platinum ELISA, eBioscience) according to the manufacturer's instructions.
Montes-Casado M., Ojeda G., Aragoneses-Fenoll L., López D., de Andrés B., Gaspar M.L., Dianzani U., Rojo J.M, & Portolés P. (2019). ICOS deficiency hampers the homeostasis, development and function of NK cells. PLoS ONE, 14(7), e0219449.
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5

Cytokine Profiling in Tumor Microenvironment

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Whole tumors, CD11b+Gr1− cells, CD90.2+ cells, CD4+ cells and CD8+ cells isolated from LLC tumors were lysed in RIPA buffer and total protein concentration was determined using a BCA Protein Assay (Pierce). Macrophage supernatants (100 μl) or 500 μg of total protein lysate from tumors were used in ELISAs to detect CCL2, TGFβ, IL-1β, TNFα, IL-6, IFNγ, IL-10, IL-12 and Granzyme B (Ready Set Go ELISA, eBioscience). Protein expression was normalized to total volume (supernatants) or mg total protein (tumor lysates).
Kaneda M.M., Messer K.S., Ralainirina N., Li H., Leem C., Gorjestani S., Woo G., Nguyen A.V., Figueiredo C.C., Foubert P., Schmid M.C., Pink M., Winkler D.G., Rausch M., Palombella V.J., Kutok J., McGovern K., Frazer K.A., Wu X., Karin M., Sasik R., Cohen E.E, & Varner J.A. (2016). PI3Kγ is a molecular switch that controls immune suppression. Nature, 539(7629), 437-442.
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6

Quantifying Inflammatory Mediators in Tissue

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Once slices were imaged, inserts were discarded and the resulting media was collected for assessment of inflammatory mediator release. Nitric oxide (NO) release was assessed by the Griess Reagent System (Promega Corporation, Madison, WI) according to the manufacturer's instructions. Briefly, samples were mixed sequentially with sulfanilamide and N-1-napthylethylenediamine dihydrochloride and incubated for 5min. Absorbance was measured at 550nm using a Wallac 1420 VICTOR plate reader (PerkinElmer, MA, USA). All samples were assayed in duplicate and nitrite content was estimated using a reference NaNO2 standard curve performed with each assay. TNF-alpha content was assessed by enzyme linked immunosorbent assay kit (ELISA; Ready-Set-Go!® ELISA, eBioscience Inc., San Diego, CA) according to the manufacturer's instructions. Briefly, samples were pipetted on 96-well plates coated with rat anti-TNF-alpha antibodies and detected using the sandwich method (anti-TNF-alpha primary antibody, avidin-HRP linked secondary antibody and tetramethylbenzidine substrate). All samples were assayed in duplicate and TNF-alpha content was estimated from a reference TNF-alpha standard curve performed with each assay.
Lutz J.A., Carter M., Fields L., Barron S, & Littleton J.M. (2015). The dietary flavonoid rhamnetin inhibits both inflammation and excitotoxicity during ethanol withdrawal in rat organotypic hippocampal slice cultures. Alcoholism, clinical and experimental research, 39(12), 2345-2353.
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7

Quantification of Inflammatory Cytokines

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Concentrations of IL-6 and TNF-α were measured by Ready-Set-Go ELISA (eBioscience, Inc., San Diego, CA). Concentrations of CCL5, CCL2, CCL3, and CXCL2 were detected by DuoSet ELISA kits from R&D Systems (Minneapolis, MN). All ELISAs were analyzed using a BioTek ELx800 (Biotek, Winooski, VT).
Fensterheim B.A., Young J.D., Luan L., Kleinbard R.R., Stothers C.L., Patil N.K., McAtee-Pereira A.G., Guo Y., Trenary I., Hernandez A., Fults J.B., Williams D.L., Sherwood E.R, & Bohannon J.K. (2018). The TLR4 agonist monophosphoryl lipid A drives broad resistance to infection via dynamic reprogramming of macrophage metabolism. Journal of immunology (Baltimore, Md. : 1950), 200(11), 3777-3789.
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8

Quantification of Cytokine Production by ELISA

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To measure IL-10 and TNF-α production, the Ready-Set-Go ELISA (Ebioscience) was used according to the manufacturer’s instructions. Briefly, plates were coated at room temperature for 3 h, after which the plates were blocked for 2 h with blocking buffer. Standards and samples (10 x diluted for TNF-α) were added to the wells and the plates incubated overnight at 4 °C. The following day, the plates were washed by filling and emptying the wells with PBS and then the biotin-conjugated antibody to mouse IL-10 and TNF-α was added for 2 h at room temperature. After washing the plates three times, streptavidin conjugated to horse radish peroxidase was added and incubated for 1 h at room temperature in the dark. After another wash, substrate was added and after allowing for proper colour development, the reaction was terminated using 2 M H2SO4. Finally, the optical density of the reaction was measured (OD450, reference filter 570 nm) using the SPECTRA MAX microplate reader (Molecular Devices) and compared with the optical density of the known standard samples to determine cytokine concentrations.
Ryan P.M., Stolte E.H., London L.E., Wells J.M., Long S.L., Joyce S.A., Gahan C.G., Fitzgerald G.F., Ross R.P., Caplice N.M, & Stanton C. (2019). Lactobacillus mucosae DPC 6426 as a bile-modifying and immunomodulatory microbe. BMC Microbiology, 19, 33.
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9

Cytokine Profiling in Cell Supernatants

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The concentrations of IL-6, monocyte chemoattractant protein-1 (MCP-1), TGF-β, IL-1β, tumor necrosis factor-alpha (TNF-α), and IL-10 were measured in cell-free supernatants using commercial ELISA kits (Ready-SET-Go! ELISA; eBio-science). Sensitivity of ELISA kits is as follows: 2 pg/mL for IL-6, 7 pg/mL for MCP-1, 8 pg/mL for TGF-β, 2.4 pg/mL for IL-1 β, 4 pg/mL, and 2 pg/mL for TNF-α and IL-10, respectively. These assays were performed in duplicates following the manufacturer’s instructions.
Alvarado-Vázquez P.A., Grosick R.L., Moracho-Vilrriales C., Ward E., Threatt T, & Romero-Sandoval E.A. (2018). Cytokine production capabilities of human primary monocyte-derived macrophages from patients with diabetes mellitus type 2 with and without diabetic peripheral neuropathy. Journal of Pain Research, 12, 69-81.
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10

Quantifying Biotinylated Chemokines

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Biotinylated chemokines (Generon, Slough) were added to media in the basal transwell chamber, incubated for 6 hours at 37°C and then quantified in apical chamber samples by “Ready-Set-Go” ELISA according to manufacturer’s instructions (eBioscience, San Diego). Streptavidin-HRP was added directly to the bound samples on the plate in the absence of secondary antibody, to detect the biotin on the bound chemokines.
Gil E., Venturini C., Stirling D., Turner C., Tezera L.B., Ercoli G., Baker T., Best K., Brown J.S, & Noursadeghi M. (2022). Pericyte derived chemokines amplify neutrophil recruitment across the cerebrovascular endothelial barrier. Frontiers in Immunology, 13, 935798.
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