Following the manufacturer’s instructions, alveolar macrophage pyroptosis was detected by the FAM-FLICA Caspase-1 Detection Kit (Immunochemistry Technologies, Minnesota, USA). The cells were gently collected and cocultured with the caspase-1 fluorescent probe in the dark at 37 ℃ for 1 h. Washing with 1X apoptosis wash buffer for removing from unbound FLICA in cells. Then, the nucleus was counterstained with Hoechst 33,342 and PI sequentially. The morphology of pyroptotic AM was identified by microscopy and analyzed with ZEN 3.1 software (Carl Zeiss).
Protocol detail
Detecting Alveolar Macrophage Pyroptosis
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Alveolar macrophage
Apoptosis
Buffer
Caspase 1
Cells
Fluorescent probe
Microscopy
Nucleus
Pyroptosis
Corresponding Organization :
Other organizations : Nanchang University, First Affiliated Hospital of Nanchang University
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Variable analysis
independent variables
- Coculture of alveolar macrophages with caspase-1 fluorescent probe
- Alveolar macrophage pyroptosis detection
- Incubation at 37 °C for 1 hour
- Washing with 1X apoptosis wash buffer
- Hoechst 33,342 and PI nuclear staining
- Positive control: Not specified
- Negative control: Not specified
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