What is the role of caspase-3?
Caspases are crucial mediators of programmed cell death (apoptosis). Among them, caspase-3 is a frequently activated death protease, catalyzing the specific cleavage of many key cellular proteins.
Where does caspase-3 activation occur?
apoptotic cell
Activation. Caspase-3 is activated in the apoptotic cell both by extrinsic (death ligand) and intrinsic (mitochondrial) pathways. The zymogen feature of caspase-3 is necessary because if unregulated, caspase activity would kill cells indiscriminately.
What activates the Wnt pathway?
The Wnt/β-catenin pathway is activated when a Wnt ligand binds to a seven-pass transmembrane Frizzled (Fz) receptor and its co-receptor, low-density lipoprotein receptor related protein 6 (LRP6) or its close relative LRP5.
Is caspase-3 Required for apoptosis?
Caspases are crucial mediators of programmed cell death (apoptosis). Caspase-3 is also required for some typical hallmarks of apoptosis, and is indispensable for apoptotic chromatin condensation and DNA fragmentation in all cell types examined.
How does caspase-3 assay work?
The Caspase-3 assay protocol is based on the formation of the chromophore p-nitroaniline (p-NA) by cleavage from the labeled substrate DEVD-pNA. Comparison of the absorbance of p-NA from an apoptotic sample with an uninduced control allows determination of the fold increase in Caspase-3 activity.
How is caspase-3 activated?
Caspase-3 is a cysteine–aspartic acid protease that cleaves cellular targets and executes cell death. Our current understanding is caspase-3 is activated by the cleavage of the interdomain linker and then subsequent cleavage of the N-terminal prodomain.
How does caspase-3 induced apoptosis?
Caspase-3 is known as an executioner caspase in apoptosis because of its role in coordinating the destruction of cellular structures such as DNA fragmentation or degradation of cytoskeletal proteins (1). The activity of caspase-3 is tightly regulated and it is produced as zymogen in an inactive pro-form (1).
How do I activate caspase-3?
How do I activate Wnt signal?
To activate Wnt signaling, one can add Wnt protein, either in a purified form or as conditioned medium to cells. Cells producing active Wnt can be obtained from the ATCC (see also the reagents page). Active Wnt protein can be obtained from several companies.
How is Wnt secreted?
In Wnt-producing cells, the Wnt protein becomes palmitoylated in the ER by the porcupine acyl-transferase. Further transport and secretion of the Wnt protein in secretory vesicles is controlled by the multi-pass transmembrane protein Wntless/Evi, which is present in the Golgi and/or on the plasma membrane.
How does caspase-3 induce apoptosis?
What does caspase-3 assay measure?
Measurement of target cell damage has historically been an important measure of CTL function. We have developed a flow cytometry-based assay for CTL-mediated cytotoxicity based on specific binding of antibody to activated caspase 3 in target cells.