Human Tumor Necrosis Factor α, TNF-α ELISA Kit

The Human Tumor Necrosis Factor Alpha (TNF-α) ELISA Kit is a highly sensitive and specific assay designed for the quantitative detection of TNF-α levels in human serum, plasma, culture media, and other biological fluids. This kit is essential for research into inflammation, immune regulation, and cancer biology.

Purpose

• Quantification of TNF-α Levels:
  • Detect and measure TNF-α concentrations in human biological samples to study inflammation, immune responses, and cancer progression.
• Sample Compatibility:
  • Suitable for serum, plasma, culture media, and other biological fluids.

Principle

The assay utilizes a Sandwich-ELISA technique for the accurate and reliable quantification of TNF-α. The process includes:

1. Pre-Coated Plate:
   • Microelisa stripplate wells are pre-coated with an antibody specific to TNF-α.
2. Sample and Standard Addition:
   • Standards and samples are added to the wells, where TNF-α in the sample binds to the immobilized antibody.
3. HRP-Conjugated Detection:
   • A Horseradish Peroxidase (HRP)-conjugated antibody specific to TNF-α is added, forming a sandwich complex.
4. Washing Step:
   • Free and unbound components are washed away to minimize background interference.
5. Substrate Reaction:
   • TMB substrate solution reacts with HRP, producing a blue color in wells containing TNF-α.
6. Stop Reaction and Measurement:
   • A stop solution is added, changing the color from blue to yellow. Optical density (OD) is measured at 450 nm using a spectrophotometer.
7. Quantification:
   • TNF-α concentrations are calculated by comparing OD values to a standard curve.

Key Features

1. High Sensitivity and Specificity:
   • Detects low levels of TNF-α with minimal cross-reactivity or interference.
2. Wide Sample Compatibility:
   • Validated for serum, plasma, culture media, and other biological fluids.
3. Quantitative Results:
   • Provides precise measurement of TNF-α concentrations using a standard curve.
4. Ease of Use:
   • Includes pre-coated plates and ready-to-use reagents for streamlined workflow.
5. Reproducibility:
   • Ensures consistent and reliable results across multiple experiments.

Kit Components

• Pre-coated Microelisa stripplate.
• TNF-α standards for standard curve generation.
• HRP-conjugated TNF-α detection antibody.
• TMB substrate solution.
• Stop solution.
• Wash buffer.
• Sample diluent.
• Instruction manual.

Applications

• Inflammation Research:
  • Study TNF-α as a key cytokine in inflammation and immune responses.
• Immune Regulation Studies:
  • Investigate TNF-α’s role in immune modulation and autoimmune disorders.
• Cancer Research:
  • Assess TNF-α levels in tumor microenvironments and its role in cancer progression and treatment resistance.
• Infectious Disease Studies:
  • Monitor TNF-α in diseases associated with systemic inflammation and immune activation.
• Therapeutic Development:
  • Evaluate TNF-α as a potential target in anti-inflammatory and cancer therapies.

Advantages

1. High Accuracy:
   • Provides precise and reproducible quantification of TNF-α.
2. Convenience:
   • Ready-to-use reagents simplify preparation and assay execution.
3. Versatility:
   • Compatible with various biological sample types.
4. Reliable:
   • Delivers consistent results for robust data interpretation.

Assay Protocol Summary

1. Add standards or samples to the pre-coated wells.
2. Incubate with HRP-conjugated TNF-α antibody.
3. Wash to remove unbound components.
4. Add TMB substrate to develop color.
5. Stop the reaction and measure OD at 450 nm.
6. Calculate TNF-α concentrations using the standard curve.

Storage and Stability

• Store all kit components at 2–8°C to maintain reagent integrity.
• Avoid repeated freeze-thaw cycles to ensure assay reliability.

The Human Tumor Necrosis Factor Alpha (TNF-α) ELISA Kit is a critical tool for researchers studying inflammation, immune responses, and cancer biology. Its precision, ease of use, and broad compatibility make it indispensable for clinical and experimental applications.