Toxicology

Apoptosis Methods in Toxicology by Perpetua M. Muganda

By Perpetua M. Muganda

This specific quantity offers a unmarried, worthy reference resource for ways that definitively determine and thoroughly quantify apoptosis. The publication starts off with universal tools applied to realize and quantitate apoptosis, in addition to apoptosis signaling pathways in toxicological and different similar examine. It keeps with multi-parametric and phased apoptosis assays for detecting early and past due apoptosis or distinguishing apoptosis from necrosis and autophagy. next chapters specialise in fresh advances in actual time and high-throughput assays that observe and quantitate apoptosis and apoptosis signaling pathways. ultimate chapters specialise in fresh advancements in preclinical anticancer therapeutics focusing on apoptosis. Written for the Methods in Pharmacology and Toxicology sequence, chapters characteristic step by step descriptions of the methodologies, in addition to professional counsel and implementation advice.

Vital and authoritative, Apoptosis equipment in Toxicology serves amateur scientists in addition to specialists, using a variety of tools from universal laboratory apparatus to high-end dear and automatic equipment in a position to acting genuine time apoptotic measurements.

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37. 38. 39. 40. 41. 42. 43. 44. 45. 46. 47. 25 Cytofluorometric detection of mitochondrial alterations in early CD95/Fas/APO-1triggered apoptosis of Jurkat T lymphoma cells. Comparison of seven mitochondrionspecific fluorochromes. Immunol Lett 61:157–163 Scaduto RC Jr, Grotyohann LW (1999) Measurement of mitochondrial membrane potential using fluorescent rhodamine derivatives. Biophys J 76:469–477 Bicknell GR, Snowden RT, Cohen GM (1994) Formation of high molecular mass DNA fragments is a marker of apoptosis in the human leukaemic cell line, U937.

Since the induction of apoptosis is concentration dependent, it is best to utilize concentrations that induce substantial apoptosis without causing necrosis. In the example experiments provided in this chapter, diepoxybutane (DEB, Sigma Cat# 202533-1G) was utilized to induce apoptosis in TK6 human lymphoblasts. 27 M stock solution; this is used to make a 1000× DEB working solution of 10 mM in sterile RPMI without serum. DEB is carcinogen and mutagen. Protective clothing and gloves should thus be worn for any work involving DEB.

When attempting to select an appropriate apoptosis detection method, it is essential to carefully analyze the purpose and the end goal of the experiment. Our lab is primarily interested in the detection of apoptosis in the context of therapies that target apoptosis signaling components to either induce or inhibit apoptosis in target cells. In this regard, it is our opinion that selectivity and early detection is critical. Also, it is important to select a platform that can distinguish between growth arrest and true cell death.

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