What is the role of caspase-3 in 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).
What is the molecular weight of caspase-3?
Alternative splicing of CPP32 results in two transcript variants which encode the same protein. The processed form of Caspase 3 consists of large (theoretical molecular weight 17kD) and small (theoretical molecular weight 12kD) subunits which associate to form an active enzyme.
Where is cleaved caspase-3?
Caspase-3 exists in the form of pro-enzyme, and is located predominantly in the cytoplasm of cells.
What is the difference between caspase-3 and cleaved caspase-3?
One specific effector caspase is caspase-3, a protein that is cleaved and thus activated upon the initiation of apoptosis. Cleaved caspase-3 propagates an apoptotic signal through enzymatic activity on downstream targets, including poly ADP ribose polymerase (PARP) and other substrates (2).
Where is caspase-3 activated?
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 is activated caspase-3?
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.
What is a limitation of the caspase-3 assay?
Additionally, the main limitations for our multiplexing protocol are that caspase-3/7 activity can determined but not quantified, and the resulting lysate cannot be used for downstream assays (i.e. Western blots or gene expression).
How does caspase cascade work?
Caspases are a family of cysteine proteases that act in concert in a cascade triggered by apoptosis signaling. The culmination of this cascade is the cleavage of a number of proteins in the cell, followed by cell disassembly, cell death, and, ultimately, the phagocytosis and removal of the cell debris.
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. The p-NA can be quantified using a spectrophotometer or a microtiter plate reader reading absorbance at 400 or 405 nm.
What is caspase 3 antibody made of?
Caspase-3 Antibody from Cell Signaling Technology. Cell Signaling Technology’s Caspase-3 Antibody is a Rabbit Polyclonal antibody. The Caspase-3 Antibody was generated using Caspase 3, and Lice as the antigen.
How do you test for activated caspase-3?
Repeat. Resuspend cells in 200-500 µl of 1X PBS and analyze on flow cytometer. Cleaved Caspase-3 (Asp175) Antibody detects endogenous levels of the large fragment (17/19 kDa) of activated caspase-3 resulting from cleavage adjacent to Asp175.
What is the difference between caspase-3 (8g10) and α-tubulin (11h10) Mab?
Caspase-3 (8G10) Rabbit mAb confirms silencing of caspase-3 expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of caspase-3 siRNA.
Where does cleavage of caspase 3 require the aspartic acid?
Cleavage of caspase-3 requires the aspartic acid residue at the P1 position (2). Fernandes-Alnemri, T. et al. (1994) J Biol Chem 269, 30761-4.