Apoptosis is the process of programmed cell death, involving a series of biochemical events leading to characteristic cell morphology changes and eventual cell death. Apoptosis is mediated by a diverse range of extracellular or intracellular signals and is important in biological phenomena such as cell termination, homeostasis, and lymphocyte interactions. Additionally, apoptotic dysregulation has been implicated in many diseases such as coronary artery disease, HIV infections, and cancer. Following TNF or Fas signaling, a critical balance between pro-apoptotic molecules (Bax, Bid, Bak, or BAD) and anti-apoptotic molecules (Bcl-xL and Bcl-2) of the Bcl-2 family is established. The pro-apoptotic homodimers interact with the mitochondrial membranes to promote the release of caspase activators such as cytochrome c and Smac. Caspase molecules are cellular cysteine proteases that perform integral steps in the apoptotic and necrotic pathways. The initiator caspases function to cleave the pro-forms of executioner caspases into their active proteolytic forms. p53, the so called “Guardian of the Genome” also plays critical role in apoptosis and is triggered by numerous cell stress events (eg, UV radiation, DNA breaks) to initiate cell death pathways. Disruption to the regulation of the p53 can result in impaired apoptosis, as is seen in some oncogenic situations. Recent reports have also shown the involvement of insulin-like growth factor binding proteins in cellular proliferations and apoptosis, namely IGFBP-3 which has anti-proliferative and pro-apoptotic properties.

RayBiotech provides a variety of apoptosis assay kits to dissect apoptotic pathways. With the RayBio® Human Apoptosis Antibody Array Kit, researchers can efficiently screen the expression levels of 43 apoptosis-related proteins. For detecting individual proteins or phosphorylated proteins, ELISA kits are available for many of the critical players in apoptosis, including p53, p27, HSP27, TRAIL, Bcl-w, BAD, cIAP-2, Smac/Diablo, and many more.

How It Works

Each capture antibody is printed onto the membrane or glass slide. Treated or untreated cell lysate samples are added to the apoptosis assay. After washing, a cocktail of biotin-conjugated anti-apoptotic antibody mix is used. Signals are visualized by chemiluminescence for membranes by incubation with HRP-Streptavidin, or by fluorescence with Cy3-Streptavidin for glass slide arrays.