Apoptosis · Antibody review
cIAP1
ABclonal · A19688
After more than a year of use, the background remains clean, with cIAP1 loss visible in several cell-death models.
01Experimental images
Lane assignments
- Lanes 1
- 0 min
- Lanes 2
- 5 min
- Lanes 3
- 15 min
- Lanes 4
- 30 min
- Lanes 5
- 60 min
- Lanes 6
- 120 min
02Observations
More than a year of use
I have used this antibody for over a year and have had good results overall. The target band is clear, with a fairly clean background and few additional bands, making changes in abundance easy to follow.
cIAP1 decreases over the time course
In this apoptosis time course, cIAP1 decreases as stimulation continues. By 2 h, accompanying caspase-3 and GSDME measurements show clear activation and cleavage, while the cIAP1 signal is declining.
Experience in other models
I have also consistently observed decreasing cIAP1 levels with this antibody in several cell-death models. For my work, the clean background and easy-to-read trend are its main advantages.
03Protein background
cIAP1, encoded by BIRC2, has a theoretical full-length mass of approximately 70 kDa and regulates cell survival, inflammation and death signaling. It acts mainly as an E3 ubiquitin ligase, controlling ubiquitination of proteins such as RIPK1 and influencing NF-κB and TNF signaling. Certain stimuli or IAP antagonists can promote cIAP1 degradation and alter cellular responses to death signals. On western blots, the full-length band is commonly used to follow its abundance and compare it with caspase or GSDME changes at the same time points.
04Experimental conditions
- Sample
- Human THP-1 monocytes
- Application
- Western blot (WB)
- Primary antibody dilution
- 1:3000
- Gel
- 4–12% gel
- Electrophoresis
- 200 V, 40 min
- Transfer
- 320 mA, 30 min
- Membrane
- 0.2 μm nitrocellulose
- Blocking
- No blocking
- Secondary antibody
- 1:5000, 1 h
- Protein loading
- 15 μL (volume; protein mass not recorded)
Comments & discussion
No sign-up. Nickname optional.