Description

Product Overview

The RayBio® Human Adiponectin ELISA Kit empowers your proteomics research with a robust, easy-to-use platform for quantifying Human Adiponectin/Acrp30. Adiponectin is a critical hormone secreted exclusively by adipose tissue, playing a vital role in regulating glucose and lipid homeostasis. As a potent insulin enhancer, it serves as a key link between adipose tissue and whole-body glucose metabolism. Understanding Adiponectin levels is essential in clinical research, as decreased levels are frequently observed in obesity and patients with coronary artery disease.

Assay Principle

This assay employs the classic sandwich ELISA technique:

1. Capture: Standards and samples are added to a 96-well plate coated with a specific anti-human Adiponectin antibody.
2. Detection: After washing, a biotinylated anti-human Adiponectin antibody is added to “sandwich” the captured Adiponectin.
3. Amplification: HRP-conjugated streptavidin is introduced, which binds to the biotinylated antibody.
4. Visualization: A TMB substrate solution is added, producing a blue color proportional to the amount of bound Adiponectin.
5. Quantification: The reaction is stopped, turning the solution yellow, and intensity is measured at 450 nm.

Kit Components & Storage

The kit is designed for convenience and longevity, with a shelf life of up to 1 year at -20°C.

• Pre-coated 96-well Microplate: 12 strips of 8 wells.
• Standard Protein: 2 vials of lyophilized Human Adiponectin.
• Detection Antibody: Biotinylated anti-Human Adiponectin.
• HRP-Streptavidin Concentrate: 80X concentrated solution.
• Assay Diluents (A, B, and C): Optimized buffers for different sample types and dilutions.
• Wash Buffer (20X): For efficient removal of unbound components.
• TMB One-Step Substrate & Stop Solution: For colorimetric detection.

Performance Characteristics

• High Sensitivity: Minimum detectable dose of 25 pg/ml.
• Excellent Recovery: Mean recovery rates range from 29% to 96.59% across serum, plasma, and cell culture media.
• Linearity: Demonstrates consistent performance with an average “percentage of expected” linearity of 92% – 95%.
• Reproducibility: Intra-assay CV < 10%; Inter-assay CV < 12%.
• Strict Specificity: No cross-reactivity with 45 tested cytokines, including TNF-alpha, IL-6, Leptin, and VEGF.

Simplified Protocol Summary

1. Prepare reagents, samples, and standards.
2. Add 100 µL standard or sample to each well; incubate 2.5 hours at RT.
3. Add 100 µL prepared biotin antibody; incubate 1 hour at RT.
4. Add 100 µL prepared Streptavidin solution; incubate 45 minutes at RT.
5. Add 100 µL TMB Substrate; incubate 30 minutes at RT.
6. Add 50 µL Stop Solution and read at 450 nm immediately.

Note: For Research Use Only. Not for use in diagnostic procedures.