Human Diacyl Glycerol(Dag) Elisa Kit, 96 Tests

The Human Diacyl Glycerol (DAG) ELISA Kit is a sensitive and specific assay designed for the quantitative measurement of DAG levels in human serum, plasma, culture media, and other biological fluids. This kit is ideal for research involving lipid signaling, metabolism, and related physiological or pathological processes.

Purpose

• Quantification of DAG Levels:
  • Measures DAG concentrations to study its role in lipid metabolism, signal transduction, and energy homeostasis.
• 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 DAG. Below is the workflow:

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

Key Features

1. High Sensitivity and Specificity:
   • Detects low levels of DAG 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 DAG 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.
• DAG standards for standard curve generation.
• HRP-conjugated DAG detection antibody.
• TMB substrate solution.
• Stop solution.
• Wash buffer.
• Sample diluent.
• Instruction manual.

Applications

• Lipid Metabolism Research:
  • Investigate DAG as a key intermediate in lipid biosynthesis and metabolism.
• Signal Transduction Studies:
  • Study DAG’s role in activating protein kinase C (PKC) and related signaling pathways.
• Energy Homeostasis Research:
  • Explore DAG as a regulator in energy storage and utilization.
• Disease Research:
  • Monitor DAG levels in conditions such as diabetes, obesity, cardiovascular disease, and metabolic disorders.
• Drug Development:
  • Assess the effects of therapeutic agents targeting lipid signaling and metabolism.

Advantages

1. High Accuracy:
   • Provides precise and reproducible quantification of DAG.
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 DAG 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 DAG 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 Diacyl Glycerol (DAG) ELISA Kit is a critical tool for researchers studying lipid signaling and metabolism. Its high sensitivity, specificity, and user-friendly design make it indispensable for lipid-related research in physiological and pathological conditions.