Related Experiment Video
Updated: Jul 5, 2026
08:51
Quantification of Coenzyme A in Cells and Tissues
Published on: September 27, 2019
An optimized luciferase bioluminescent assay for coenzyme A
Simone M Marques1, Joaquim C G Esteves da Silva
1Centro de Investigação em Química (CIQ-UP), Chemistry Department, Faculty of Sciences, University of Porto, R. Campo Alegre 687, 4169-007, Porto, Portugal.
Analytical and Bioanalytical Chemistry
|April 26, 2008
Summary
A novel bioluminescent assay quantifies coenzyme A (CoA) using firefly luciferase. This optimized method provides a sensitive and robust tool for CoA analysis, with a linear response and reliable detection limits.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Enzymology
Background:
- Coenzyme A (CoA) is a vital molecule in cellular metabolism.
- Accurate quantification of CoA is essential for understanding metabolic pathways.
- Existing CoA quantification methods may lack sensitivity or robustness.
Purpose of the Study:
- To develop a novel bioluminescent method for quantifying coenzyme A (CoA).
- To optimize the assay parameters for enhanced sensitivity and reliability.
- To establish the performance characteristics of the new CoA quantification assay.
Main Methods:
- Enzymatic conversion of dehydroluciferyl-adenylate (L-AMP) to dehydroluciferyl-coenzyme A (L-CoA) using firefly luciferase (LUC).
- Bioluminescent light emission measured using a luminometer.
- Optimization of L-AMP, luciferase, ATP, and luciferin concentrations via experimental design.
Main Results:
- Linear response for CoA quantification observed in the range of 0.25-4 microM.
- Limit of detection (LOD) established at 0.24 microM.
- Limit of quantification (LOQ) determined to be 0.80 microM.
- Assay demonstrated a relative standard deviation of approximately 7%.
Conclusions:
- The developed bioluminescent method offers a sensitive and robust approach for CoA analysis.
- The optimized assay provides reliable quantification within a defined concentration range.
- This method is suitable for various applications requiring accurate CoA measurement.
