Caspases detection techniques

Comparison of Techniques for Detection of Caspases to TUNEL Method 10... 

Both techniques can be applied to most experimental models of apoptosis, including cells in culture and biopsies. In addition, techniques based on the cleavage of synthetic caspase substrates can be applied to caspases, activated in vitro and in vivo [82, 83], Whereas Western blotting is time consuming and mainly a qualitative assay, the detection of caspase activity by cleavage of synthetic substrates is a quantitative, relatively fast and sensitive method [82]. However, both techniques do not provide information about the type and distribution of the cells with activated caspases in the tissue examined. 

Immunohistochemistry, on the other hand, enables identification of activated caspases or their cleaved products in fixed archival tissue sections. This technique allows identification of cell(s) undergoing caspase activation, as well as analysis of the distribution of cell(s) in the tissue. Specific antibodies to various caspases are now commercially available, the most frequently studied being caspase-3. Studies in various human tissues and cells have shown that immunohistochemical detection of activated caspase-3 is a useful tool for identifying apoptotic cells in archival material, even before all of the morphological features of apoptosis occur [84-86]. Several target proteins cleaved by caspases can also be detected by immunohistochemistry for example PARP [87], actin [88, 89], and lamin B [90]. 

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