Apoptosis · Antibody review
BCL2
Beyotime · AG1225
After about three years of use, target bands remain clear and the background clean; BCL2 is fairly stable in this experiment.
01Experimental images
02Observations
An antibody I have used for three years
I have used this BCL2 antibody for about three years, and it still performs very well. The target band is distinct, the background clean and there are hardly any noticeable extra bands.
BCL2 remains stable at 2 h
By 2 h, accompanying caspase-3 and GSDME measurements show clear activation and cleavage, while total BCL2 remains fairly stable. There is no obvious decrease here, an informative contrast alongside the other apoptosis-related changes.
03Protein background
BCL2 is an anti-apoptotic BCL2-family protein whose full-length band is usually near 26 kDa. Through interactions with pro-apoptotic family members, it limits mitochondrial outer-membrane permeabilization and cytochrome c release, helping cells resist apoptotic stimuli. Its function depends on protein abundance as well as binding partners and modifications such as phosphorylation. Total BCL2 may therefore change little after a short stimulation period, while longer time courses or different treatments can produce different expression patterns.
04Experimental conditions
- Sample
- Human THP-1 monocytes
- Application
- Western blot (WB)
- Primary antibody dilution
- 1:3000
- Gel
- 4–12% gel
- Electrophoresis
- 200 V, 35 min
- Transfer
- 300 mA, 30 min
- Membrane
- 0.2 μm nitrocellulose
- Blocking
- 1% PVP, 1 h
- Secondary antibody
- 1:8000, 1 h
- Protein loading
- Not recorded
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