The Mouse Tumor Necrosis Factor-alpha (TNF-α) ELISA Kit is designed for the quantitative detection and measurement of mouse TNF-α in biological fluids such as serum, plasma, and cell culture supernatants. TNF-α is primarily produced by activated T cells and macrophages in response to inflammation or infection, but is also produced by B cells, NK cells, mast cells, neutrophils, dendritic cells, microglia, endothelial cells, smooth muscle cells, cardiomyocytes, and fibroblasts. TNF-α is a pro-inflammatory cytokine and stimulates anti-tumor responses in vitro.
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: 15.6 - 2000 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 5 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 88/532.
• Recovery: A mid-curve recovery of 84 - 86% 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 2.4% (CV). The Inter-assay precision of this assay is 3.7% (CV).
* National Institute of Biological Standards and Control, Potters Bar, Hertfordshire EN6 3QG, UK.
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
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Mouse TNF-alpha ELISA Kit
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Mouse TNF-alpha ELISA Kit
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Mouse TNF-alpha ELISA Kit
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Mouse TNF-alpha ELISA Kit
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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 B Cell Research
Workflow Stages for Dendritic Cell Research
Workflow Stages for Granulocyte Research
Workflow Stages for Monocyte Research
Workflow Stages for Natural Killer Cell Research
Workflow Stages for T Cell Research
Workflow Stages for Cardiac and Skeletal Muscle Research
Resources and Publications
Educational Materials (2)
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