2-Color Immunocytochemistry of Cytochrome C and Mitochondria
The MSA07 Apoptosis Detection Kit contains an anti-cytochrome C monoclonal antibody (50 ug) and an isotype-specific secondary antibody (50ug) which has a fluorescein fluorophore conjugated to it. Apoptotic cells which have released mitochondrial cytochrome C into the cytosol can be differentiated from non apoptotic cells which still retain cytochrome c in their mitochondria by fluorescence microscopy. The kit also contains an anti-Complex V α monoclonal antibody (50 ug) and the appropriate isotype-specific secondary antibody (50ug) which has a Texas Red fluorophore conjugated to it. Complex V is an inner mitochondrial membrane protein and serves as a mitochondrial marker since it is not released into the cytoplasm during apoptosis, unlike cytochrome c. Thus one is able to observe cytochrome c and a mitochondrial marker (Complex V alpha) simultaneously by fluorescence microscopy using MSA07. Buffers and blocking solution for immunocytochemical analysis are also included in the kit.
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