Canine Cytochrome-C (Cyt-C) ELISA Kit

The Canine Cytochrome-C (Cyt-C) ELISA Kit is a highly sensitive and specific assay designed for the quantitative measurement of Cytochrome-C (Cyt-C) in canine serum, plasma, culture media, and other biological fluids. This kit uses a Sandwich-ELISA method to deliver accurate and reliable results, making it ideal for research applications.

Purpose

• Quantification of Canine Cytochrome-C:
  • Measure Cyt-C levels in a variety of biological samples for research into mitochondrial function, apoptosis, and oxidative stress.
• Applicable Samples:
  • Serum, plasma, culture media, and other biological fluids from canine sources.

Principle

The assay employs a Sandwich-ELISA technique for high specificity and sensitivity. The steps include:

1. Pre-coated Plate:
   • The Microelisa strip plate is pre-coated with an antibody specific to canine Cyt-C.
2. Sample and Standard Addition:
   • Standards and samples are added to the wells, allowing Cyt-C in the sample to bind to the immobilized antibody.
3. HRP-Conjugated Detection:
   • A Horseradish Peroxidase (HRP)-conjugated antibody specific for Cyt-C is added, forming a sandwich complex with Cyt-C.
4. Washing Step:
   • Unbound components are washed away, reducing background noise.
5. TMB Substrate Reaction:
   • A TMB substrate is added, producing a blue color in the presence of HRP.
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 correlates directly with Cyt-C concentration.

Features

1. High Sensitivity and Specificity:
   • Detects low levels of Cyt-C with minimal cross-reactivity.
2. Quantitative Results:
   • Generates a standard curve for precise quantification of Cyt-C.
3. Wide Sample Compatibility:
   • Suitable for canine serum, plasma, and culture media.
4. Ready-to-Use Components:
   • Pre-coated plates and all necessary reagents included.
5. Reproducibility:
   • Ensures consistent results across experiments.

Kit Components

• Pre-coated Microelisa strip plate.
• Standards for Cyt-C quantification.
• HRP-conjugated antibody specific for Cyt-C.
• TMB substrate solution.
• Stop solution.
• Wash buffer.
• Sample diluent.
• Instruction manual.

Applications

• Mitochondrial Research:
  • Study mitochondrial dysfunction and the role of Cyt-C in apoptosis.
• Oxidative Stress Studies:
  • Quantify Cyt-C levels as a marker of oxidative stress in canine samples.
• Veterinary Research:
  • Investigate Cyt-C as a biomarker for various diseases in canines.
• Cell Death Mechanisms:
  • Explore the role of Cyt-C in apoptosis and cell death pathways.

Assay Protocol Summary

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

Storage and Stability

• Store kit at 2–8°C to maintain reagent stability.
• Ensure all reagents are equilibrated to room temperature before use.

Advantages

• Ease of Use:
  • User-friendly protocol with pre-coated plates and ready-to-use reagents.
• High Precision:
  • Accurate quantification of Cyt-C levels in canine samples.
• Versatile Applications:
  • Suitable for research in apoptosis, mitochondrial function, and oxidative stress.

The Canine Cytochrome-C (Cyt-C) ELISA Kit offers a reliable and efficient method for measuring Cyt-C in various canine biological samples. Its sensitivity, specificity, and straightforward protocol make it an invaluable tool for research in mitochondrial health, apoptosis, and oxidative stress.