Canine Kidney injury molecule 1 (Kim-1) ELISA Kit

The Canine Kidney Injury Molecule 1 (Kim-1) ELISA Kit is a highly sensitive and specific assay designed for the quantitative measurement of Kim-1 levels in canine serum, plasma, culture media, and other biological fluids. This kit utilizes a Sandwich-ELISA technique to ensure accurate and reproducible results, making it ideal for research into kidney function, injury, and related diseases in canines.

Purpose

• Quantification of Canine Kim-1:
  • Measures the concentration of Kim-1 as a biomarker for kidney injury and function in canines.
• Applicable Samples:
  • Canine serum, plasma, culture media, and other biological fluids.

Principle

The assay is based on the Sandwich-ELISA technique, which ensures high specificity and sensitivity for Kim-1 detection. The workflow includes:

1. Pre-Coated Plate:
   • Microelisa strip plate wells are pre-coated with an antibody specific to canine Kim-1.
2. Sample and Standard Addition:
   • Standards and samples are added to the wells, allowing Kim-1 in the sample to bind to the immobilized antibody.
3. HRP-Conjugated Detection:
   • A Horseradish Peroxidase (HRP)-conjugated antibody specific for Kim-1 is added, forming a sandwich complex.
4. Washing Step:
   • Unbound components are washed away, reducing background interference.
5. TMB Substrate Reaction:
   • TMB substrate is added, which reacts with the HRP enzyme, producing a blue color in positive wells.
6. Stop Reaction:
   • A stop solution is added, converting 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 directly proportional to the Kim-1 concentration in the sample.

Features

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

Kit Components

• Pre-coated Microelisa strip plate.
• Standards for generating a standard curve.
• HRP-conjugated Kim-1 detection antibody.
• TMB substrate solution.
• Stop solution.
• Wash buffer.
• Sample diluent.
• Instruction manual.

Applications

• Renal Disease Research:
  • Study the role of Kim-1 as a biomarker for acute and chronic kidney injury in canines.
• Veterinary Diagnostics:
  • Monitor kidney health and diagnose renal disorders in dogs.
• Toxicology Studies:
  • Assess nephrotoxicity in drug development or environmental exposures.
• Inflammation and Repair Studies:
  • Investigate Kim-1's role in renal inflammation, damage, and repair mechanisms.

Advantages

1. Accuracy:
   • High specificity for Kim-1 ensures minimal interference and precise quantification.
2. Convenience:
   • Ready-to-use reagents simplify the workflow.
3. Versatility:
   • Can be used for various sample types and experimental designs.
4. Reproducibility:
   • Ensures reliable results across multiple tests.

Assay Protocol Summary

1. Add standards or samples to the pre-coated wells.
2. Incubate with HRP-conjugated Kim-1 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 Kim-1 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 Kidney Injury Molecule 1 (Kim-1) ELISA Kit provides a reliable and efficient method for measuring Kim-1 levels in canine biological samples. Its high sensitivity, specificity, and ease of use make it an invaluable tool for veterinary research, renal health monitoring, and toxicology studies.