Pyroptosis · Antibody review
Caspase-1 p20
ABclonal · A23429
After about three years, p20 is still easy to detect; the inhibitor groups also show an interesting intermediate-sized band.
01Experimental images
Lane assignments
- Lanes 1, 2, 8, 9, 11, 12
- Not recorded
- Lanes 3, 4
- Stimulation (drug not specified)
- Lanes 5, 6
- Inhibitor added (not specified)
- Lanes 7, 10
- Apoptotic stimulation (drug not specified)
02Observations
Still detecting p20 after about three years
I bought this antibody about three years ago and still find p20 fairly easy to detect. Stimulated lanes 3 and 4 differ clearly from the controls, with a prominent low-molecular-weight p20 band.
An intermediate-sized band with inhibitors
With inhibitor treatment, lanes 5 and 6 show a clear band between 30 and 40 kDa. Its position suggests a possible p33-like intermediate accumulating during caspase-1 processing, which I found interesting.
Multiple bands after apoptotic stimulation
Apoptotic stimulation in lanes 7 and 10 produces bands at several molecular weights. Alongside the usual p20 signal, this antibody shows a broader pattern of processing-related changes that is worth examining together.
03Protein background
Caspase-1 is usually present as the approximately 45–46 kDa precursor pro-caspase-1. After inflammasome assembly, the precursor is recruited, dimerizes and undergoes self-cleavage to form active p33/p10. Further cleavage removes the CARD domain and releases p20/p10 from the inflammasome; in cells, this release can reduce its activity. Caspase-1 also processes IL-1β, IL-18 and GSDMD. Western blots may therefore show full-length p45/46 together with fragments such as p33, p20 and p10, depending on the antibody epitope, cell type and sampling time.
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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