Cells were harvested, transferred to 96-well plates (2 × 105 cells per well), and stimulated in vitro with 100 ng/mL lipopolysaccharide (LPS; Sigma, St Louis, MO, USA) for 24 h at 37°C and 5% CO2. Supernatants were collected and stored at -80°C until further analysis. Concentrations of cytokines, including interferon (IFN)-γ, interleukin (IL)-10, and IL-4, in culture supernatants were measured with ELISA MAX sets (BioLegend, San Diego, CA, USA), according to the manufacturer’s instructions. All samples were analyzed in triplicate.
Elisa max sets
Manufactured by BioLegend
Sourced in United States
ELISA MAX sets are a collection of reagents and components designed for the performance of Enzyme-Linked Immunosorbent Assay (ELISA) experiments. These sets include pre-coated capture antibodies, detection antibodies, standards, and necessary buffers to facilitate the ELISA process. The core function of ELISA MAX sets is to provide researchers with a comprehensive and streamlined solution for measuring the concentration of specific proteins or analytes in biological samples.
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Lab products found in correlation
4 protocols using elisa max sets
1
Cytokine Secretion Assay for LPS-Stimulated Cells
2
Serum Interleukin-1β Quantification
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Blood samples were collected by cardiac puncture under anesthesia. For serum preparation, blood was allowed to clot for 30 minutes at room temperature followed by centrifugation (2000g) for 20 minutes at 4°C. Serum was collected and stored at −20°C. Quantification of interleukin‐1β in serum samples was performed using ELISA MAX Sets (BioLegend, San Diego, CA).
3
Cytokine Profiling by ELISA
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The amounts of TNF-α, IL-1β, IL-10, IL-6, IL-8, GM-CSF, and transforming growth factor beta (TGF-β) were determined in control-CM, H37Rv-CM, and M-CM by ELISA, according to manufacturer's instructions (TNF-α, IL-1β, IL-8, and TGF-β, eBioscience, San Diego, CA, USA; IL-10, IL-6, and GM-CSF, ELISA MAX™ Sets BioLegend, San Diego, CA, USA).
4
Optimizing Ovalbumin Vaccination in Mice
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After anesthetization, mice were administered 3 μg of OVA in PBS (60 μL) containing 3, 10, or 30% (w/v) HP-β-CyD and K3 CpG-ODN (0.1, 1, or 10 μg) at the base of the tail on days 0 and 14. Sera were collected at 7 days after the boost injection. Then, the mice were euthanized and their spleens were collected. After hemolysis of red blood cells using ACK lysis buffer, splenocytes were cultured at a density of 1 × 107 cells/mL in RPMI medium containing penicillin (100 units/mL)/streptomycin (100 μg/mL) and 10% fetal bovine serum, and stimulated with OVA (10 μg/mL) for 48 h. The concentrations of mouse IL-4, IL-5, and IFN-γ were measured using ELISA MAX™ sets in accordance with the manufacturer's instructions (Biolegend, San Diego, CA, USA).
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