Ready set go elisa kit

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Germany, France

Ready-Set-Go ELISA kits are a line of products designed for enzyme-linked immunosorbent assay (ELISA) applications. These kits provide pre-coated microplates, standards, and reagents necessary to perform ELISAs. The core function of these kits is to facilitate the detection and quantification of specific analytes in sample matrices.

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Lab products found in correlation

253 protocols using ready set go elisa kit

Quantification of Cytokine Secretion and Receptor Levels

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To determine IFN-γ and IL-17 secretion, cells isolated from CNS draining lymph nodes were adjusted to 5 × 10⁶ cells (c)/ml in RPMI 1640 medium, containing 8.6% FCS, 4 mM glutamine, 1 mM sodium pyruvate, 0.1 mM MEM non-essential amino acids, 200 μg/ml penicillin, 200 μg/ml streptomycin, 25 μg/ml gentamicin, and 50 μM 2-ME and stimulated with MOG_35–55 (MEVGWYRSPFSRVVHLYRNGK) (Ana Spec, Fremont, CA) in Costar® 96-well round-bottom culture plates (Thermo Fisher Scientific Inc., Waltham, MA) for 96 h. Cell-free supernatants were collected and cytokines were quantified by ELISA using Ready-SET-Go!® ELISA kits (eBioscience, San Diego, CA). To determine TNF, TNFR1, and TNFR2 concentrations, serum samples were collected and quantified using eBioscience Ready-SET-Go!^® ELISA kits (TNF and TNFR2) or the mouse TNFR1 DuoSet^® ELISA kit (R&D Systems, Minneapolis, MN). All cell-free culture supernatants and serum samples were stored at −80°C until use. The absorbance (450 nm) minus background (570 nm) of the colorimetric reactions were quantified using a Synergy^™ 4 spectrophotometry microplate reader (BioTek, Winooski, VT). The lower limit of sensitivity of the TNF assay was 8 pg/ml. The recognition of TNF by the Ready-SET-Go!^® assay is not affected by 1000-fold excess soluble TNFR1 or TNFR2.

Miller P.G., Bonn M.B., Franklin C.L., Ericsson A.C, & McKarns S.C. (2015). TNFR2 Deficiency Acts in Concert with Gut Microbiota to Precipitate Spontaneous Sex-biased CNS Demyelinating Autoimmune Disease. Journal of immunology (Baltimore, Md. : 1950), 195(10), 4668-4684.

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Cytokine Secretion and Gene Expression in Colon Explants

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Colon explants were cultured for 24 h (37 °C, 5% CO₂) to measure secretion of cytokines IL-9, IL-13 and IL-22 in the supernatants using mouse uncoated ELISA kits (Invitrogen, Ready-SET-Go! ELISA kits, eBioscience), or for 4 h in the presence of 200 ng/ml IL-36α to measure Muc2 mRNA expression. IL-6 and TNF-α were measured in the serum of mice after HFD using mouse uncoated ELISA kits (Invitrogen, Ready-SET-Go! ELISA kits, eBioscience). Levels of fasting plasma insulin were measured using Ultra Sensitive Mouse Insulin ELISA Kit (Crystal Chem) according to the manufacturer’s instructions.

Giannoudaki E., Hernandez-Santana Y.E., Mulfaul K., Doyle S.L., Hams E., Fallon P.G., Mat A., O’Shea D., Kopf M., Hogan A.E, & Walsh P.T. (2019). Interleukin-36 cytokines alter the intestinal microbiome and can protect against obesity and metabolic dysfunction. Nature Communications, 10, 4003.

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T Cell Response to OVA-loaded DCs

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2×10⁵ OT-1 CD8⁺ T cells were co-cultured with 2×10⁴ OVA_257-264-loaded mDCs in 96-well plates with or without the same number of liver LSed-DCs. 48 h later, IL-2, IFN-γ, and IL-10 levels in co-culture supernatants were determined using the Ready-SET-Go ELISA Kit (eBioscience, San Diego, CA, USA) according to the manufacturer’s instructions. For NO detection, mDCs and LSed-DCs were cultured for 24 h with or without LPS (0.5 μg/mL). NO production was determined by measuring nitrite concentration using the Griess assay as described previously [43 (link)].

Wang Q., He H., Chen D., Wang C., Xu Y, & Song W. (2017). Hepatic stroma-educated regulatory DCs suppress CD8+ T cell proliferation in mice. Oncotarget, 8(55), 93414-93425.

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Quantification of Cytokine Levels in Mesothelioma

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The concentrations of cytokines within conditioned media following culture with T cells, mesothelioma cell lines (1 × 10⁶/mL) or sorted tumor cells were quantified using a competitive enzyme-linked immunoassay according to the manufacturers’ instructions. The following molecules were tested GM-CSF (Biolegend), IL13 (BD Biosciences), IL8 (BioLegend), IL6 (BioLegend), G-CSF (R&D Systems), VEGF (R&D Systems), Mesothelin (BioLegend). The concentration of IFNγ in coculture supernatants was determined by Ready Set Go ELISA kit (eBioscience).

Khanna S., Graef S., Mussai F., Thomas A., Wali N., Yenidunya B.G., Yuan C., Morrow B., Zhang J., Korangy F., Greten T.F., Steinberg S.M., Stetler-Stevenson M., Middleton G., De Santo C, & Hassan R. (2018). Tumor-Derived GM-CSF Promotes Granulocyte Immunosuppression in Mesothelioma Patients. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(12), 2859-2872.

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Cytokine Profiling in THP-1 and Synoviocytes

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The THP-1 cells (5 × 10⁴ cells per well of a 96-well plate) were incubated with CM. After a 48-h incubation, the supernatants were collected, followed by the determination of human IL-8, IL-10, IL-12, and TNF-α using the Ready-Set-Go ELISA kit (eBioscience, San Diego, CA, USA) according to the manufacturer’s instructions. The synoviocytes (5 × 10⁴ cells per well of a 96-well plate) were treated with IL-1β and HA. After treatment for 48 h, the supernatants were collected, followed by the determination of human IL-6 and PGE2 assay kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Lee C.H., Chiang C.F., Kuo F.C., Su S.C., Huang C.L., Liu J.S., Lu C.H., Hsieh C.H., Wang C.C., Lee C.H, & Shen P.H. (2021). High-Molecular-Weight Hyaluronic Acid Inhibits IL-1β-Induced Synovial Inflammation and Macrophage Polarization through the GRP78-NF-κB Signaling Pathway. International Journal of Molecular Sciences, 22(21), 11917.

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Generating Secondary Effector T Cells

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Thy1.1⁺ OT-I cells (1.0 × 10⁴) were transferred i.v. and mice were infected with 10⁴ CFU Listeria monocytogenes strains expressing OVA APL epitope (LM-OVA APLs) (15 (link)) 24 h later. Secondary effectors were generated in 4 week post-infection mice by immunizing with 50 µg N4 OVA peptide (GenScript) emulsified in IFA (Gibco) in both hind foot pads. Five days later, draining popliteal LNs were collected and pooled for analysis. Serum was analyzed using Ready-Set-Go ELISA kit according to manufacturer’s instructions (eBiosciences).

Krummey S.M., Chen C.W., Guasch S.A., Liu D., Wagener M., Larsen C.P, & Ford M.L. (2016). Enhanced Requirement for TNFR2 in Graft Rejection Mediated by Low Affinity Memory CD8+ T Cells During Heterologous Immunity. Journal of immunology (Baltimore, Md. : 1950), 197(5), 2009-2015.

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Plasma Lipid and Inflammation Analysis

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Mice were fasted overnight and at the time of necropsy, blood was collected by cardiac puncture and plasma was frozen at −80°C until analyzed. Total plasma cholesterol and triglyceride levels were determined using the Cobas c311 automated chemistry analyzer, with reagents, calibrators, and controls from Roche Diagnostics. Plasma LPS levels were determined using Pierce LAL Chromogenic Endotoxin Kit (Thermo Fisher) and IL‐6 levels were measured by Ready‐set‐go ELISA kit from eBiosciences. Aliquots of undiluted plasma were also used to determine total IAP activity as described above.

Ghosh S.S., He H., Wang J., Korzun W., Yannie P.J, & Ghosh S. (2018). Intestine‐specific expression of human chimeric intestinal alkaline phosphatase attenuates Western diet‐induced barrier dysfunction and glucose intolerance. Physiological Reports, 6(14), e13790.

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Investigating IL-10-Dependent B Cell Functions

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IL-10-GFP mice were injected IV on D0, D2, and D5 with 20 × 10⁶ apoptotic thymocytes. On D7 mice were sacrificed and blood, peritoneal lavage and spleens harvested. Splenic CD19^+ve B cells were FACs sorted into IL-10-GFP^{+ve or −ve} fractions, phenotyped and cultured for 10 days in the presence of MegaAPRIL [Adipogen (200 ng/ml)], CpGB [ODN1826 Eurofins MWG Operon (1 μg/ml)], and IL-4 [R&D System (50 ng/ml)], after which culture supernatants were checked for IgM and IgG levels (Ready-SET-Go ELISA kit eBioScience).

Miles K., Simpson J., Brown S., Cowan G., Gray D, & Gray M. (2018). Immune Tolerance to Apoptotic Self Is Mediated Primarily by Regulatory B1a Cells. Frontiers in Immunology, 8, 1952.

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GM-CSF Expression Assessment

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GM-CSF level was assayed by Ready-SET-Go ELISA kit (eBioscience) according to the manufactures' instructions.

Sheng W., Yang F., Zhou Y., Yang H., Low P.Y., Kemeny D.M., Tan P., Moh A., Kaplan M.H., Zhang Y, & Fu X.Y. (2014). STAT5 programs a distinct subset of GM-CSF-producing T helper cells that is essential for autoimmune neuroinflammation. Cell Research, 24(12), 1387-1402.

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Measuring IFNγ in DC-CD4 T-cell Cocultures

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DCs and CD4 T-cells were sorted as described above. 2500 – 3000 DCs were then co-cultured with 100,000 CD4 T-cells in 120 µL of volume in a 384 well culture plate. After 60 hours of co-culture at 37 degrees C, supernatant was assessed for IFNγ using the Ready-Set-Go ELISA kit (eBioscience, San Diego, CA) according to manufacturer’s instructions. Where noted, ex vivo cultures were supplemented with 1 × 10⁵ PFU of inactivated HSV-2.

Soerens A.G., Da Costa A, & Lund J.M. (2016). Regulatory T-cells are essential to promote proper CD4 T-cell priming upon mucosal infection. Mucosal immunology, 9(6), 1395-1406.

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