Canine Neutrophil Gelatinase Associated Lipocalin (NGAL) ELISA Kit

The Canine Neutrophil Gelatinase Associated Lipocalin (NGAL) ELISA Kit is a sensitive and specific assay designed for the quantitative measurement of NGAL levels in canine serum, plasma, culture media, or other biological fluids. Utilizing a Sandwich-ELISA technique, this kit delivers accurate and reproducible results, making it ideal for research in renal function, inflammation, and related conditions in canines.

Purpose

• Quantification of Canine NGAL:
  • Measures NGAL concentrations to study kidney function, injury, and inflammatory responses in dogs.
• Applicable Samples:
  • Canine serum, plasma, culture media, and other biological fluids.

Principle

The kit employs the Sandwich-ELISA technique for reliable detection and quantification of NGAL. The workflow is as follows:

1. Pre-Coated Plate:
   • Microelisa strip plate wells are pre-coated with an antibody specific to canine NGAL.
2. Sample and Standard Addition:
   • Standards and samples are added to the wells, allowing NGAL in the sample to bind to the immobilized antibody.
3. HRP-Conjugated Detection:
   • A Horseradish Peroxidase (HRP)-conjugated antibody specific for NGAL is added, forming a sandwich complex with the captured NGAL.
4. Washing Step:
   • Unbound components are washed away to minimize background noise.
5. TMB Substrate Reaction:
   • TMB substrate is added, reacting with the HRP enzyme to produce a blue color in positive wells.
6. Stop Reaction:
   • A stop solution is added, turning the color from blue to yellow.
7. Optical Density Measurement:
   • The optical density (OD) is measured at 450 nm using a spectrophotometer. The OD value is proportional to NGAL concentration in the sample.

Features

1. High Sensitivity and Specificity:
   • Detects low levels of NGAL with minimal cross-reactivity.
2. Quantitative Results:
   • Generates a standard curve for precise measurement of NGAL concentrations.
3. Wide Sample Compatibility:
   • Suitable for a variety of biological fluids, including serum, plasma, and culture media.
4. Ease of Use:
   • Ready-to-use reagents and pre-coated plates simplify assay setup.
5. Reproducibility:
   • Consistent results across experiments due to stringent quality control.

Kit Components

• Pre-coated Microelisa strip plate.
• Standards for generating a standard curve.
• HRP-conjugated NGAL detection antibody.
• TMB substrate solution.
• Stop solution.
• Wash buffer.
• Sample diluent.
• Instruction manual.

Applications

• Renal Function Research:
  • Study NGAL as a biomarker for kidney injury, particularly acute kidney injury (AKI) in canines.
• Inflammatory Response Studies:
  • Investigate NGAL’s role in neutrophil activity and inflammation.
• Veterinary Diagnostics:
  • Diagnose and monitor kidney disorders or inflammatory diseases in dogs.
• Toxicology Research:
  • Assess renal toxicity in drug development or environmental exposure studies.

Advantages

1. Accuracy:
   • High specificity ensures precise measurement of NGAL levels with minimal interference.
2. Convenience:
   • Pre-coated plates and ready-to-use reagents reduce preparation time.
3. Versatility:
   • Can be used for various sample types and experimental designs.
4. Reproducibility:
   • Reliable results across multiple tests.

Assay Protocol Summary

1. Add standards or samples to the pre-coated wells.
2. Incubate with HRP-conjugated NGAL 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 NGAL concentration using the standard curve.

Storage and Stability

• Store kit at 2–8°C for long-term stability.
• Avoid repeated freeze-thaw cycles of reagents to maintain quality.

The Canine Neutrophil Gelatinase Associated Lipocalin (NGAL) ELISA Kit offers a reliable and efficient method for measuring NGAL levels in canine biological samples. Its high sensitivity, specificity, and straightforward protocol make it an invaluable tool for veterinary research, renal function studies, and inflammation monitoring.