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Updated: Jan 17, 2026
Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages
Published on: May 21, 2018
Protocol for quantifying extracellular and intracellular ATP from macrophages upon inflammasome activation using a
Julieta Schachter1, Adriana Guijarro2, Pablo J Schwarzbaum1
1Department of Biological Chemistry and Institute of Biochemistry and Biophysics (IQUIFIB), School of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina.
Abstract:
Extracellular ATP triggers several cellular responses, while intracellular ATP depletion primes cells for death. Here, we present a protocol for the quantification of extracellular and intracellular ATP from bone marrow-derived macrophages upon inflammasome activation using a luciferin-luciferase technique. We describe steps for macrophage isolation, differentiation, and inflammasome activation with real-time ATP measurement. The protocol allows quantitative determination of intracellular ATP and extracellular ATP hydrolysis, with applicability to different cell types. For complete details on the use and execution of this protocol, please refer to Schachter et al.1.
Insights
This study details a method to measure extracellular and intracellular ATP in macrophages during inflammasome activation. The protocol quantifies ATP levels and hydrolysis, aiding cell death research.
Area of Science:
- Cellular Biology
- Immunology
- Biochemistry
Background:
- Extracellular ATP (adenosine triphosphate) influences cellular functions.
- Intracellular ATP depletion is linked to programmed cell death.
- Inflammasome activation is a critical immune pathway.
Purpose of the Study:
- To present a protocol for quantifying extracellular and intracellular ATP.
- To enable real-time ATP measurement during inflammasome activation in macrophages.
- To provide a method applicable to various cell types.
Main Methods:
- Isolation and differentiation of bone marrow-derived macrophages.
- Induction of inflammasome activation.
- Quantification of extracellular and intracellular ATP using a luciferin-luciferase assay.
- Real-time ATP measurement.
Main Results:
- Successful quantification of intracellular ATP levels.
- Measurement of extracellular ATP hydrolysis.
- Demonstration of the protocol's applicability to different cell types.
Conclusions:
- The developed protocol enables precise measurement of ATP dynamics.
- This method is valuable for studying inflammasome activation and cell death.
- The protocol offers a versatile tool for cellular ATP analysis.
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