Pyroptosis · Antibody review
GSDME
ABclonal · A28230
The full-length band drops markedly while p35 NT is clear, making the processing pattern easy to see.
01Experimental images
02Observations
A result I was happy with
This antibody gave a visibly good result. After apoptotic stimulation, lanes 7 and 10 show a very prominent NT band at approximately 35 kDa.
Reading full-length and NT together
As the p35 band appears, the full-length GSDME band falls markedly and becomes barely visible. Seeing full-length loss alongside NT accumulation makes the processing pattern easy to read.
03Protein background
GSDME, also called DFNA5, is a gasdermin-family protein with a full-length size usually near 55 kDa. It connects apoptotic signaling with membrane lysis. Caspase-3 cleaves its linker region to release an approximately 30–35 kDa N-terminal fragment, which can form membrane pores and cause pyroptotic lysis in GSDME-expressing cells. Western blots commonly follow loss of the full-length band and accumulation of the N-terminal band. Its apparent migration, intensity and timing vary with cell type, treatment conditions and the region recognized by the antibody.
04Experimental conditions
- Sample
- Human THP-1 monocytes
- Application
- Western blot (WB)
- Primary antibody dilution
- 1:3000
- Gel
- 4–20% gradient gel
- Electrophoresis
- 200 V, 45 min
- Transfer
- 320 mA, 30 min
- Membrane
- 0.2 μm nitrocellulose
- Blocking
- No blocking
- Secondary antibody
- 1:5000, 1 h
- Protein loading
- Not recorded
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