Scientist has used Cleaved Caspase 3 to determine apoptosis. Also they have used Caspase3/7 assay for apoptosis. I am wondering if in "Caspase3/7 assay", the Caspase 3 and Caspase 7 are cleaved. Why Caspase 3 and 7 are assayed as combined assay as Caspase 3/7. Please clarify this confusion to me?
The Caspase-Glo® 3/7 Assay(a,b) is a homogeneous, luminescent assay that measures caspase-3 and -7 activities. These members of the cysteine aspartic acid-specific protease (caspase) family play key effector roles in apoptosis in mammalian cells. However, the sensitivity of caspase3/7 assay kit is very low. That is why I always check the expression level of active caspase 3 and cleaved PARP.
https://www.promega.com/-/media/files/resources/protocols/technical-bulletins/101/caspase-glo-3-7-assay-protocol.pdf
Interesting question ...but answer is very simple. The assay provides a luminogenic caspase-3/7 substrate, which contains the tetrapeptide sequence DEVD, in a reagent optimized for caspase activity, luciferase activity and cell lysis. caspase-3 is often the primary source of cellular "DEVDase" activity, other "effector" caspases, such as caspase-7 may also contribute to cleavage of DEVD sequence present in proteins. ex PARP...it can be cleaved by both 3 and 7. so one can not say its just due to 3 or 7 its 3/7 activity.
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