Assays and reagents to examine the structure, function, and physiology of apoptosis
Cell apoptosis, also known as programmed cell death, is a highly regulated process that not only allows for proper growth and development by ridding the organism of unneeded cells and tissues, but also minimizes threats to the organism by destroying surplus cells of the immune system and virus-infected or DNA-damaged cells .
Programmed cell death is morphologically and biochemically distinct from cell death by injury (necrosis). Unlike necrotic cells, apoptotic cells exhibit compaction of the nuclear chromatin, shrinkage of the cytoplasm, and production of membrane-bound apoptotic bodies, as well as DNA fragmentation and cleavage or degradation of several cellular proteins (Figure 1). Biochemically, apoptosis is distinguished by fragmentation of the genome and cleavage or degradation of several cellular proteins. Incorrectly regulated apoptosis is implicated in a number of disease states, including cancer, stroke, Alzheimer’s disease, and several autoimmune diseases.
Apoptosis can be induced through three different pathways: 1) targeting mitochondria functionality (mitochondrial, cellular or apoptosis intrinsic pathway), 2) direct transduction of the signal via adaptor proteins (death-receptor or apoptosis extrinsic pathway) and 3) the perforin/granzyme pathway [2-3].
Figure 1. Comparison of apoptosis and necrosis.
Identifying cells that are going through apoptosis can be challenging because many of the assays used as indicators are detecting structural and functional changes that occur in other processes as well. Additionally, no single parameter fully defines programmed cell death in all systems and the appearance of these changes can vary with apoptotic pathway or cell types. Therefore, it is often advantageous to use several different assays to detect cell apoptosis.
Interested in the non-apoptotic form of cell death? Learn more about Ferroptosis Research Solutions.
The characteristic breakdown of the nucleus during apoptosis includes collapse and fragmentation of the DNA, degradation of the nuclear envelope and nuclear blebbing, resulting in the formation of micronuclei.
Changes in the cell’s membrane include distortion of the cell membrane, changes in the lipid composition and membrane integrity, and translocation of the phospholipid phosphatidylserine.
Apoptosis is a highly regulated process and involves a significant number of signaling pathways. Thermo Fisher Scientific offers a wide range of primary antibodies, ELISA kits, multiplexed immunoassays, peptides and recombinant proteins as well as genomic assays targeted to specific proteins. Below is just an example of the types of assays we have available for some key apoptosis proteins.
Table 2. Example of products for key apoptosis pathway proteins.
|Protein||Full name||Primary antibodies||ELISA Kits||Proteins/peptides||TaqMan® assays|
|AIF||Apoptosis inducing factor||AIF antibodies||AIF proteins/peptides||AIF TaqMan® Assays|
|AIF-1/IBA1||Allograft inflammatory factor 1||AIF-1/IBA1 antibodies||AIF-1/IBA1 TaqMan® Assays|
|Apaf-1||Apoptotic protease activating factor||Apaf-1 antibodies||Apaf-1 TaqMan® Assays|
|Bcl Protein Family|
(Bcl-2, Bcl-10, Bcl-x, Bcl-xL, Bcl-xS, BCL-w)
|B-cell lymphoma protein||Bcl Protein Family antibodies||Bcl Protein Family ELISAs||Bcl Protein Family proteins/peptides||Bcl Protein Family TaqMan® Assays|
|t-BID/BID-p15||Truncated BH3-interacting domain death agonist; BH3-interacting domain death agonist p15||t-BID/BID-p15 antibodies||BID ELISAs||t-BID/BID-p15 TaqMan® Assays|
|BIM/BCL2L11||Bcl-2-like protein 11||BIM antibodies||BIM proteins||BIM TaqMan® Assays|
|BAD||Bcl2-associated agonist of cell death||BAD antibodies||BAD TaqMan® Assays|
|BAK||Bcl-2 homologous antagonist/killer||BAK antibodies||BAK TaqMan® Assays|
|BNIP3||BCL2/adenovirus E1B 19 kDa protein-interacting protein 3||BNIP3 antibodies||BNIP3 TaqMan® Assays|
|Caspase-3||Cysteinyl aspartic acid-protease-3||Caspase-3 antibodies||Caspase-3 ELISAs||Caspase-3 proteins/peptides||Caspase-3 TaqMan® Assays|
|Caspase-7||Cysteinyl aspartic acid-protease-7||Caspase-7 antibodies||Caspase-7 ELISAs||Caspase-7 proteins/peptides||Caspase-7 TaqMan® Assays|
|Caspase-9||Cysteinyl aspartic acid-protease-9||Caspase-9 antibodies||Caspase-9 ELISAs||Caspase-9 proteins/peptides||Caspase-9 TaqMan® Assays|
|cIAP1/BIRC2||Cellular inhibitor of apoptosis protein 1; Baculoviral IAP repeat-containing protein 2||cIAP1/BIRC2||cIAP1 ELISAs 1||cIAP1 proteins/peptides||cIAP1 TaqMan® Assays|
|cIAP2/BIRC3||Cellular inhibitor of apoptosis protein 2; Baculoviral IAP repeat-containing protein 3||cIAP2/BIRC3||cIAP2 TaqMan® Assays|
|Cytochrome c||Cytochrome c antibodies||Cytochrome c ELISAs||Cytochrome c TaqMan® Assays|
|FADD||FAS-associated death domain protein||FADD TaqMan® Assays|
|Fas/CD95||Fas/CD95 antibodies||Fas/CD95 ELISAs||Fas/CD95 proteins/peptides||Fas/CD95 TaqMan® Assays|
|Fas-ligand/CD178||Fas-ligand/CD178 antibodies||Fas-ligand/CD178 ELISAs||Fas-ligand/CD178 TaqMan® Assays|
|MCL-1||Myeloid cell leukemia-1||MCL-1 ELISAs||MCL-1 proteins/peptides||MCL-1 TaqMan® Assays|
|Smac/DIABLO||Second mitochondrial activator of caspases/direct||Smac/DIABLO antibodies||Smac/DIABLO ELISAs||Smac/DIABLO proteins/peptides||Smac/DIABLO TaqMan® Assays|
Cell Analysis Support Center
Find technical information, tips and tricks, and answers to everyday problems.
For Research Use Only. Not for use in diagnostic procedures.