C57BL/6 mice were treated with DMXAA or vehicle control and serum was obtained 6 hours later. ELISA was performed for IFN-β, TNF-α and IL-6 using pre-coated plates (Biolegend) according to the manufacturer protocol.
Pre coated plates
Manufactured by BioLegend
Sourced in United States
Pre-coated plates are a type of laboratory equipment used for various applications in biomedical research and diagnostics. These plates have a pre-applied coating, which can be specific to the intended use, such as protein, antibody, or cell-based assays. The pre-coating helps to streamline the experimental setup and provides a consistent and standardized surface for the desired application.
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Lab products found in correlation
Legend max human il 33 elisa kit, by BioLegend (1 mentions)
Legend max free active tgf β1 elisa kit, by BioLegend (1 mentions)
Legend max human il 6 elisa kit, by BioLegend (1 mentions)
Legend max total tgf β1 elisa kit, by BioLegend (1 mentions)
Elisa max deluxe, by BioLegend (1 mentions)
Elisa max standard set, by BioLegend (1 mentions)
Multiskan ex microplate reader, by Thermo Fisher Scientific (1 mentions)
8 protocols using pre coated plates
1
DMXAA-induced Cytokine Response
2
Cytokine Profiling in Tumor-Bearing Mice
3
Serum IgE and IL-33 Quantification
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Blood samples were obtained from the patients who did not receive IT, the patients who received IT for 6 months and for 2 years and from the healthy controls were left at room temperature for 30 minutes until they coagulated. They were then centrifuged at 2,000 rpm for 10 minutes until serums were obtained. The serum samples were stored at −70 C until the study. The total IgE and IL-33 were determined in the serum samples. The total IgE levels were measured by enzyme linked immunosorbent assay (ELISA) according to the Ridascreen A0141 kit manufacturer's instructions (R-Biopharm AG, Darmstadt, Germany). Interleukin-33 was determined by ELISA according to the kit manufacturer's instructions, LEGEND MAX Human IL-33 ELISA Kit with pre-coated plates (Biolegend, San Diego, USA). The minimal detection level of IL-33 was 4.14 pg/mL.
4
Measuring Active TGF-β1 Levels in Patients
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Active TGF-β1 serum levels in patients and healthy donors were measured using LEGEND MAX Free Active TGF-β1 ELISA Kit with pre-coated plates (Biolegend). The assay was run according to the manufacturer’s recommendations.
5
IL6 Quantification in Caco-2 Cells
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To quantify protein expression, media was collected from Caco-2 cells at 12 and 24 h post treatment and stored at −80 °C. IL6 content of media samples was determined via LEGEND MAX™ Human IL6 ELISA kit with pre-coated plates (Biolegend, San Diego, Ca.) according to manufacturer’s protocol.
6
Quantification of Cytokine Levels in Melanoma and Macrophage Samples
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CM from melanoma cells or supernatants from macrophages were used for sandwich ELISA experiments using commercially available kits (see also the table for reagents above). 96-well NUNC clear flat-bottom plates were coated with antibodies (IL-10, IL-8, IL-1α and TNF-α) and incubated O/N at 4°C in a humified chamber. ELISA MAX™ standard sets (Biolegend) were used for the detection of IL-10, IL-8 and TNF-α; ELISA MAX Deluxe (Biolegend) for IL1-α; and for TGF-β, LEGEND MAX Total TGF-β1 ELISA Kit with pre-coated plates was used (Biolegend). Microwell absorbance was read at 450 nm on a Thermo Scientific Multiskan EX microplate reader. All samples were quantified based on a standard curve using Microsoft Excel.
7
Multiplex Cytokine Immunoassay Protocol
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Multiplex fluorescent bead-based immunoassay with Diac-loneDIAplex kit (Gen-Probe, Besanc ¸on, France) was used for IL-17A and IFN-g measurements. IL-9 concentration was measured by ELISA using precoated plates (Biolegend).
8
Th17 Cell Differentiation and IL-17A Quantification
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Splenic naïve CD4 + T cells were cultured in Th17 polarizing conditions and then re-stimulated with coated anti-mouse CD3e mAb (1 µg/ml) (BD Biosciences PharMingen, San Diego, CA, USA) and soluble anti-mouse CD28 mAb (2 µg/ml) (BD Biosciences PharMingen) for 40 h. The mice were humanely euthanized and mouse serum was obtained at day 6 of IMQ treatment. Mouse serum and culture supernatant IL-17A levels were determined using the LEGEND MAX™ mouse IL-17A enzyme-linked immunosorbent assay (ELISA) kit with pre-coated plates (BioLegend, Inc, San Diego, CA, USA), according to the manufacturer's instructions.
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