The major executers in the apoptotic progam are proteases know as caspases (cysteine-dependent, aspartate- specific proteases). Caspase family orchestrate the changes of the cell during apoptotic process. Caspases exist as precursor, which, when activated iniciate the death program by destroying cellular key components and activating factors that mediate damage to the cells. A cleavage in the caspase 3 precursor induces it to activate and leads to the activated caspase-3 form. Our Caspase 3-7 Activity Assay Cell Line allows to monitor the caspase-3 activity level through the turboFP602 protein distribution in cell-based assays.
Caspase 3-7 Activity Assay Cell Line contains a reporter polypeptide which comprises the next parts:
- A mitocondrial localization signal (MTS) which localizes the protein in the mitochondria
- The Red Fluorescent Protein FP602
- The caspase-3 cleavage sequence DEVD, which is the link between previous peptides
In non-apoptotic cells, the tFP602 appears localized in mitochondria; in a perinuclear localization due to de MTS situated in the N-terminal of the construct. After promoting apoptosis, caspases are activated and cleave the specific caspase-clavage site (CCS) situated between the MTS and the tFP602 in the construct, resulting in liberation of tFP602 and entry into the nucleus. So, you can measure apoptosis by image analysis, quantifying the red fluorecence intensity inside the cell nucleus.
