Assay Name
In Vitro Mitochondrial Membrane Potential-based Apoptosis Assay (FCM)
Short Description
C1498-cell based In Vitro Mitochondrial Membrane Potential-based Apoptosis Assay (FCM)
Assay Description
During apoptosis, the mitochondrion membrane potential (ΔΨm) decreases and the changes in membrane potential can be detected by FCM using different fluorescence dyes. Dyes such as chloro-methyl-X rhodamine, 3,3'-dihexyloxacarbocyanine and rhodamine 123 are commonly utilized dyes to evaluate the mitochondrial membrane potential). These dyes can penetrate into mitochondria and remain until the membrane potential decreases and move into the cytoplasm when the ΔΨm decreases. JC-1 (cyanine dye) is another example, whose fluorescence changes from green to red (red fluorescence when the dye is concentrated and green fluorescence when it is released). The ratio between green to red fluorescence can be measured by FCM to detect changes in ΔΨm. Tetramethylrhodamine perchlorate and ethyl ester are other examples of dyes behaving similar to JC-1.
Assay Type
Detection of Apoptosis Assays
Assay Type Details
Apoptosis (programmed cell death) plays a vital role in embryonic development, homeostasis, functioning of immune system and wound repair. The ability to evade induction of apoptosis has been used by cancer cells to survive against host defense mechanisms. The molecular mechanisms involved in cancer cell apoptosis have been well documented and it involves certain biochemical events such as DNA fragmentation, chromatin condensation, cell organelle degradation and protein cleavage, etc. The extrinsic and intrinsic (mitochondrial) pathways are the two major pathways involved in apoptosis. With the available techniques and assays, a number of apoptosis inducing agents (natural compounds, synthetic compounds, nano-formulations, peptides and enzymes) in many cancer cells have been identified. Selection of an assay for apoptosis detection is based on factors such as apoptotic pathway, nature of drug, cell type being used and the method of analysis.
Assay Subtype
Flow Cytometry to Detect Apoptosis Assays
Assay MOA
Flow Cytometry Assay