Cell Signaling Technology
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb
#9664
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Our aim was to stain epithelial tissue biopsy samples (fixed in formalin and embedded in paraffin) with caspase-3 to estimate the levels of apoptotic cells in the epithelium. We selected this product because it has been used in the literature and because the images on the supplier's website looked fairly convincing.
Immunohistochemistry
Formalin-fixed paraffin-embedded epithelial biopsies
1:1000, for 1 hour at room temperature
TBST/5% Normal Goat Serum (as recommended)
SignalStain® Boost IHC Detection Reagents (as recommended)
None
SignalStain® DAB Substrate Kit (as recommended)
The results were largely negative on our epithelial biopsy samples; however, we always included a "positive control" section from tonsil. Staining in the tonsil was not spectacular, but there were clear positive cells (see image).
It works!
Needed a fair amount of optimization (especially antigen retreival)
Make sure you always include a positive control! You will get reliable results with this antibody, but it may take a few attempts.