Human Zonulin ELISA Kit

The Human Zonulin ELISA Kit is a highly sensitive and specific assay designed for the quantitative detection of Zonulin levels in human serum, plasma, culture media, and other biological fluids. This kit is essential for research into intestinal permeability, gut health, and related physiological or pathological processes.

Purpose

• Quantification of Zonulin Levels:
  • Detect and measure Zonulin concentrations in human biological samples to study gut barrier function, intestinal permeability, and related disorders.
• Sample Compatibility:
  • Suitable for serum, plasma, culture media, and other biological fluids.

Principle

The assay employs a Sandwich-ELISA technique for accurate and reliable quantification of Zonulin. The process includes:

1. Pre-Coated Plate:
   • Microelisa stripplate wells are pre-coated with an antibody specific to Zonulin.
2. Sample and Standard Addition:
   • Standards and samples are added to the wells, where Zonulin in the sample binds to the immobilized antibody.
3. HRP-Conjugated Detection:
   • A Horseradish Peroxidase (HRP)-conjugated antibody specific to Zonulin 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 Zonulin.
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:
   • Zonulin concentrations are calculated by comparing OD values to a standard curve.

Key Features

1. High Sensitivity and Specificity:
   • Detects low levels of Zonulin 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 Zonulin 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.
• Zonulin standards for standard curve generation.
• HRP-conjugated Zonulin detection antibody.
• TMB substrate solution.
• Stop solution.
• Wash buffer.
• Sample diluent.
• Instruction manual.

Applications

• Gut Health Research:
  • Study Zonulin as a biomarker for intestinal permeability and gut barrier integrity.
• Autoimmune Disorder Studies:
  • Investigate Zonulin levels in diseases like celiac disease and type 1 diabetes.
• Metabolic Research:
  • Explore Zonulin’s role in obesity, metabolic syndrome, and related disorders.
• Clinical Diagnostics Research:
  • Monitor Zonulin as a biomarker for leaky gut syndrome and gastrointestinal conditions.
• Therapeutic Development:
  • Evaluate treatments targeting gut health and intestinal barrier restoration.

Advantages

1. High Accuracy:
   • Provides precise and reproducible quantification of Zonulin.
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 Zonulin 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 Zonulin 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 Zonulin ELISA Kit is a vital tool for researchers studying gut health, intestinal permeability, and related disorders. Its precision, ease of use, and broad compatibility make it indispensable for clinical and experimental applications.