Cell-based assays provide a wealth of information and are valuable tools in drug discovery applications. Reducing high attrition rates due to toxicity in drug development continues to be a key challenge
for the pharmaceutical industry.
Cell viability, proliferation, and apoptosis are generally considered together
despite the fact that they measure different parameters.
Most in vitro proliferation assays determine the number of
viable cells and are based on quantifying the number of cell nuclei,
metabolic activity, assessing cell redox potential or mitochondrial integrity.
It is important to understand which cellular event a researcher plans to
measure in order to accurately address the overall aims and objective.
Below is a compilation of commonly used assays to assist in correct selection for a specific experimental design. These proliferation assays can be multiplexed with cytotoxicity or apoptosis assays that monitor
cell fate in response to compound treatment.
Compound-induced cell apoptosis can be measured by an increase in cell membrane permeability, loss of membrane asymmetry or caspases activation.
Determine the ratio of live and dead cells. The parameters that define cell viability can be as diverse as the redox potential of the cell population, the integrity of cell membranes, or the activity of cellular enzymes. Cell viability assays can determine the effect of drug candidates on cells and used to optimize cell culture conditions.
Assess the number of live and dead cells in a population following treatment with a drug candidate or pharmacological agent under investigation.
Cell proliferation is the biological process of cells increasing in number over time through cell division. Cell proliferation plays a vital role in regular tissue and cellular homoeostasis for proper growth, development, and maintenance of organism. Proliferation assays monitor the growth rate of cell populations.
(Programmed Cell Death type 1) is essential for development, homeostasis, and in the pathogenesis of various diseases such as cancer. Apoptotic cells in a controlled fashion occur in response to extrinsic or intrinsic signals. The hall mark of apoptotic cell death include exposure of phosphatidylserine on the extracellular face of the plasma membrane, activation of caspases, disruption of mitochondrial membrane potential, cell shrinkage, DNA fragmentation and DNA condensation.
(Programmed Cell Death type 2) is selective degradation of intracellular targets, such as misfolded proteins and damaged organelles, thereby serving as an important homeostatic function. Autophagy in essence is a biological recycling mechanism where misfolded proteins are ubiquitinated and targeted for degradation by lysosomal pathway.
(Programmed Cell Death type 3) is characterized by cell swelling and destruction of the plasma membrane and subcellular organelles. Necrotic cell death is considered a heterogeneous phenomenon including both programmed and accidental cell death.
We offer assays to assess cell health, such as cell senescence assays that detect senescence markers associated with β-galactosidase activity which reflects cell membrane integrity.
Kinases are a large and diverse group of enzymes which are the focus of roughly 1/3 of all drug development efforts due to their association with multiple diseases. Consequently, there is demand for assays that can detect kinase activity and subsequently their modulation by drug compounds. This has led to multiple commercial offerings that employ a variety of different detection technologies. Many of these assays employ microplates to allow for the measurement of 96 up to 1536 samples in parallel for which a multi-mode microplate reader is employed to read the experiment.
Kinase binding assays
Kinase activity assays based on ATP-ADP conversion
Kinase activity assays based on substrate conversion
Substrate phosphorylation assays