The Human Interleukin 1 beta (IL-1β) ELISA Kit is designed for the quantitative detection and measurement of human IL-1β in biological fluids such as serum, plasma, and cell culture supernatants. IL-1β is secreted by monocytes, macrophages, and dendritic cells. IL-1β is a proinflammatory cytokine that recruits immunocompetent cells to affected sites by stimulating the expression of adhesion molecules.
The assay is based on the sandwich ELISA method, in which samples are added to ELISA strip plates pre-coated with capture antibodies specific for the cytokine. The captured cytokine is detected by addition of a biotinylated detection antibody, followed by streptavidin-horseradish peroxidase, which binds the biotinylated antibody. Addition of the chromogenic enzyme substrate 3,3’,5,5’ tetramethylbenzidine (TMB) results in a colored product with an intensity directly proportional to the concentration of cytokine in the sample. The concentration of the cytokine is determined by comparison to a serial dilution of the cytokine standard analyzed in parallel.
The assay is based on the sandwich ELISA method, in which samples are added to ELISA strip plates pre-coated with capture antibodies specific for the cytokine. The captured cytokine is detected by addition of a biotinylated detection antibody, followed by streptavidin-horseradish peroxidase, which binds the biotinylated antibody. Addition of the chromogenic enzyme substrate 3,3’,5,5’ tetramethylbenzidine (TMB) results in a colored product with an intensity directly proportional to the concentration of cytokine in the sample. The concentration of the cytokine is determined by comparison to a serial dilution of the cytokine standard analyzed in parallel.
Data Figures
Representative Standard Curve
• Reportable Range: 1 - 316 pg/mL. This is the concentration range in which measurement of the analyte can be done with the highest precision, accuracy, and linearity.
• Sensitivity: The limit of detection of this assay is 4 pg/mL. This is the analyte concentration with absorbance two standard deviations higher than the zero standard.
• Accuracy: The analyte standard of this ELISA was calibrated against NIBSC international standard 86/680.
• Recovery: A mid-curve recovery of 83 - 98% was determined by spiking defined amounts of analyte standard into serum or plasma samples in repeated experiments.
• Precision: The intra-assay precision of this assay is 2.6% (CV). The inter-assay precision of this assay is 3.6% (CV).
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
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Applications
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Research Area
Workflow Stages
Workflow Stages for Dendritic Cell Research
Resources and Publications
Educational Materials (2)
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