Product Description
The BioDeviceLab Human Tumor Necrosis Factor Alpha (TNF-α) ELISA Kit is a high-performance enzyme-linked immunosorbent assay developed for the quantitative determination of TNF-α in biological samples. TNF-α is a central pro-inflammatory cytokine produced by immune cells and epithelial cells in response to microbial products, tissue injury, and inflammatory stimuli.
TNF-α plays a key role in regulating epithelial tight junction integrity, immune cell recruitment, apoptosis, and inflammatory amplification in the gastrointestinal tract. Elevated TNF-α levels are strongly associated with inflammatory bowel disease, infection, dysbiosis, and epithelial barrier dysfunction. In gut-on-chip platforms, TNF-α serves as a critical biomarker for quantifying inflammatory barrier disruption, immune–epithelial crosstalk, and therapeutic response to anti-inflammatory or biologic agents. This assay combines high analytical sensitivity, strong specificity, and robust reproducibility, making it suitable for both mechanistic inflammation studies and advanced biodevice validation workflows.
Intended Use
This kit is intended for the quantitative determination of human TNF-α in:
• Serum
• Plasma
• Cell culture supernatants
• Tissue culture media
Applications include gut-on-chip inflammation modeling, epithelial barrier disruption studies, host–microbiome immune interaction research, inflammatory bowel disease research, and biodevice calibration and benchmarking.
Assay Principle
This assay employs a sandwich ELISA format:
• TNF-α present in the sample binds to capture antibodies immobilized on a 96-well microplate.
• A biotinylated detection antibody specific to TNF-α binds to the captured protein.
• An enzyme conjugate is added, followed by chromogenic substrate (TMB).
• The enzymatic reaction generates a colorimetric signal measured at 450 nm.
• Absorbance is directly proportional to the TNF-α concentration in the sample.
Calibration Curve
• A calibration curve is generated using recombinant human TNF-α standards with known concentrations.
• Mean absorbance values are plotted against TNF-α concentration on a semi-logarithmic scale.
• A four-parameter logistic (4PL) regression model is recommended for curve fitting.
• Sample concentrations are calculated by interpolation from the standard curve.
Analytical Performance
Dynamic detection range: 5 – 3,000 pg/mL
Limit of detection (LOD): ≤ 2 pg/mL
Intra-assay CV: < 5%
Inter-assay CV: < 5%
Selectivity & Specificity
The assay demonstrates high specificity for TNF-α with minimal cross-reactivity to other cytokines (IL-1β, IL-6, IL-8, IFN-γ) (< 1%).
Kit Components
Each kit contains sufficient reagents for up to 96 determinations:
• Antibody-coated 96-well microplate
• Recombinant human TNF-α standards
• Biotinylated detection antibody
• Enzyme conjugate
• Assay buffer
• Wash buffer (10×)
• TMB substrate
• Stop solution
• Detailed assay protocol
Required Materials (Not Provided)
• Microplate reader capable of measuring absorbance at 450 nm
• Adjustable micropipettes and disposable tips
• Plate washer or manual washing equipment
• Plate shaker (recommended)
• Deionized or distilled water
Sample Collection & Handling
• Collect samples using standard laboratory procedures.
• Avoid repeated freeze–thaw cycles.
• Samples exceeding the highest standard should be diluted using assay buffer.
• Particulate matter should be removed prior to analysis.
Quality Control
• Standards and controls should be included in each assay run.
• Duplicate measurements are recommended.
• Acceptance criteria should be established according to experimental requirements.
BioDeviceLab applies quality-controlled reagent preparation and batch-to-batch consistency practices to support reproducibility and scalability for biodevice development.
Storage & Stability
• Store all components at 2–8 °C.
• Do not freeze reagents unless explicitly stated.
• Unopened kits are stable until the stated expiration date.
• Opened reagents should be handled according to protocol recommendations.
Limitations
• For research use only.
• Not intended for diagnostic or therapeutic procedures.
• Results should be interpreted within the context of experimental design and inflammatory model.
Applications
• Gut-on-chip inflammatory barrier disruption studies
• Host–microbiome immune crosstalk research
• Inflammatory bowel disease modeling
• Drug and biologic screening
• Biodevice calibration and benchmarking
