Human Glycated Hemoglobin A1c(HbA1c) ELISA Kit

The Human Hemoglobin A1C (HbA1C) ELISA Kit is a highly sensitive and specific assay designed for the quantitative detection of HbA1C levels in human serum, plasma, culture media, and other biological fluids. This kit is ideal for research on diabetes management, glucose metabolism, and related disorders.

Purpose

• Quantification of HbA1C Levels:
  • Measures HbA1C concentrations as a biomarker for long-term blood glucose control.
• Sample Compatibility:
  • Suitable for human serum, plasma, and other biological fluids.

Principle

The assay employs a Sandwich-ELISA technique for the specific and accurate measurement of HbA1C. Below is the workflow:

1. Pre-Coated Plate:
   • Microelisa stripplate wells are pre-coated with an antibody specific to HbA1C.
2. Sample and Standard Addition:
   • Standards and samples are added to the wells, allowing HbA1C in the sample to bind to the immobilized antibody.
3. HRP-Conjugated Detection:
   • A Horseradish Peroxidase (HRP)-conjugated antibody specific to HbA1C 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 with HbA1C.
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:
   • HbA1C concentrations are calculated by comparing OD values to a standard curve.

Key Features

1. High Sensitivity and Specificity:
   • Accurately detects low levels of HbA1C with minimal cross-reactivity or interference.
2. Wide Sample Compatibility:
   • Suitable for serum, plasma, culture media, and other biological fluids.
3. Quantitative Results:
   • Provides precise measurement of HbA1C concentrations using a standard curve.
4. Ease of Use:
   • Includes pre-coated plates and ready-to-use reagents for a streamlined workflow.
5. Reproducibility:
   • Ensures consistent and reliable results across multiple experiments.

Kit Components

• Pre-coated Microelisa stripplate.
• HbA1C standards for standard curve generation.
• HRP-conjugated HbA1C detection antibody.
• TMB substrate solution.
• Stop solution.
• Wash buffer.
• Sample diluent.
• Instruction manual.

Applications

• Diabetes Research:
  • Monitor HbA1C levels as a biomarker for long-term blood glucose control.
• Metabolic Studies:
  • Investigate the relationship between HbA1C and glucose metabolism.
• Endocrine Research:
  • Study HbA1C in relation to insulin resistance and hormonal regulation.
• Drug Development:
  • Evaluate therapeutic interventions targeting glucose control.
• Clinical Diagnostics Research:
  • Support diagnostics for diabetes and glucose-related disorders.

Advantages

1. High Accuracy:
   • Provides precise and reproducible quantification of HbA1C.
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 HbA1C 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 HbA1C 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 Hemoglobin A1C (HbA1C) ELISA Kit is a vital tool for research in diabetes management and glucose metabolism. Its high sensitivity, specificity, and user-friendly design make it indispensable for studying long-term glycemic control and related disorders.