Guava Technologies has released an application poster titled 'Detection of Compounds Inducing Distinct Caspase Pathways'.
It describes how researchers used Guava Caspase reagents to identify a unique compound from an extensive panel that differentially induced the intrinsic pathway of caspase activation in HeLa but not Jurkat cells.
Apoptosis, or programmed cell death, is characterised by a progressive series of morphological and biochemical changes that result in fragmentation of DNA and disassembly of the cell proper.
Initiation of the apoptotic cascade is a tightly regulated process complicated by the myriad of signalling pathways contributing to its control.
The ability to monitor mechanisms of apoptotic induction is critical in drug discovery for designing potent yet precisely targeted therapeutic compounds.
The transduction of apoptotic signals through initiator caspase (caspase 8 or 9) is stimulated through independent triggering mechanisms and is thus subject to unique control.
Guava's application poster describes how researchers used a Guava Easycyte Plus System and the Guava Caspase Software Module to test a series of 165 cytotoxic, immunosuppressive, anti-proliferative and anti-inflammatory compounds for their ability to induce apoptosis using non-adherent human T cells (Jurkat) and adherent human cervical cancer cells (HeLa).
The application poster demonstrates how combining proprietary assays enables the elucidation of whether one or both pathways is primarily induced and what the potency of the various compounds is for inducing a particular pathway.
The poster demonstrates how screening multiple markers of apoptotic induction with Guava's cellular assay kits enables a rapid and more complete understanding of the mechanisms of action of compounds.
Guava Caspase 8 and 9 Assays employ a Flica (fluorescent labelled inhibitor of caspases) reagent, which contains an amino acid peptide inhibitor sequence targeted for either active caspase 8 or 9.
Guava Caspase 8 and 9 Assays and Flica reagents work with adherent or suspended cells using tube or 96-well plate protocols.
They are non-cytotoxic and cell permanent, eliminating the need for cell lysis and permeabilisation procedures.
Guava Caspase 8 and 9 Assays provide direct and absolute cell counts and population percentages, with staining and data acquisition possible in less than three hours
