Molecular cell biology for drug discovery





Biological testing is an integral part of drug discovery process. The range of in vitro molecular biology tests we perform includes:
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Cytometric analysis of apoptosis and necrosis (AnxV/PI staining)

Apoptosis is a physiological process of controlled cell death that is essential during embryonic development and in maintenance of tissue homeostasis. New drugs associated with apoptosis are expected to be most effective against tumors with high proliferation rates and are being screened for use in the treatment of cancer. The best known apoptotic cells surface marker is phosphatidylserine (PS). Exposure of PS on the surface of dying cells allows safe and efficient clearance of apoptotic waste thus preventing induction of inflammation. Biocentrum offers high-sensitive and precise method of cell death identification with the use of fluorescently labeled annexin V (AnxV) that specifically binds to PS with high affinity. Additional use of propidium iodide (PI) and professional flow cytometry analysis enables to characterize type of cell death (apoptotic or necrotic) with high reliability. Analysis of apoptosis/necrosis together with cell cycle arrest identification constitute excellent complement to cytotoxicity research.

Cytometric analysis of cell cycle inhibition (PI staining)

Studies of the proliferation characteristics of normal and malignant cells provide broad spectrum of information about molecular mechanisms underlying cell replication, understanding mechanisms of action of drugs and designing of effective cancer therapy strategies. Cell cycle arrest is the phenomenon that plays a key role in therapy of cancer, AIDS and cardiovascular disease. A lot of naturally occurring and synthetic molecules regarded as potential anti-cancer drugs affect basal cell cycle control mechanisms and modulate cell cycle events thus having potential roles in cancer therapeutics and prevention. Cell cycle arrest analysis similarly with apoptosis identification and cytotoxicity assays becomes basic method used in anti-cancer drug discovery and development pipeline. The most commonly used dye for DNA content and cell cycle analysis is propidium iodide (PI) that intercalates into the major groove of double-stranded DNA and produces a highly fluorescent adduct. The principle of the method is to stain fixed and ribonuclease treated cells with PI and cytometric analysis in aim to characterize number of cells residing in each of three analyzed cell phases (G0/G1, S and G2/M).