By Perpetua M. Muganda
This special quantity offers a unmarried, beneficial reference resource for tactics that definitively determine and safely quantify apoptosis. The e-book starts with universal tools applied to realize and quantitate apoptosis, in addition to apoptosis signaling pathways in toxicological and different similar learn. It keeps with multi-parametric and phased apoptosis assays for detecting early and overdue apoptosis or distinguishing apoptosis from necrosis and autophagy. next chapters specialize in contemporary advances in genuine time and high-throughput assays that realize and quantitate apoptosis and apoptosis signaling pathways. ultimate chapters specialise in contemporary advancements in preclinical anticancer therapeutics focusing on apoptosis. Written for the Methods in Pharmacology and Toxicology sequence, chapters function step by step descriptions of the methodologies, in addition to specialist counsel and implementation advice.
Vital and authoritative, Apoptosis tools in Toxicology serves beginner scientists in addition to specialists, using quite a number tools from universal laboratory gear to high-end pricey and automatic equipment in a position to acting genuine time apoptotic measurements.
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Extra resources for Apoptosis Methods in Toxicology
The most common method utilized to determine the extent of phosphatidylserine (PS) externalization in apoptotic cells is the annexin V-FITC assay . This assay is performed on live cells using fluorescence microscopy or flow cytometry. Most of the annexin-V-FITC assay kits on the market include a cell impermeable DNA binding fluorescent dye, such as propidium iodide (PI). This enables the distinction of live cells (with intact membranes, and thus PI negative) and necrotic cells (with damaged membranes, and thus PI positive).
Chondria in CCCP-induced apoptosis. Exp Cell Res 299:533–540 Terauchi S, Yamamoto T, Yamashita K, Kataoka M, Terada H, Shinohara Y (2005) Molecular basis of morphological changes in mitochondrial membrane accompanying induction of permeability transition, as revealed by immuno-electron microscopy. Mitochondrion 5:248–254 Galluzzi L, Zamzami N, de La Motte RT, Lemaire C, Brenner C, Kroemer G (2007) Methods for the assessment of mitochondrial membrane permeabilization in apoptosis. Apoptosis 12:803–813 Loeffler M, Daugas E, Susin SA, Zamzami N, Metivier D, Nieminen AL et al (2001) Dominant cell death induction by extramitochondrially targeted apoptosis-inducing factor.
In one recent promising example, [18F]4-fluorobenzylcarbonyl–Val–Ala– Asp(OMe)–fluoromethylketone was developed and validated in vivo as a novel PET imaging probe capable of labeling active caspases and detecting apoptotic cell death triggered by anticancer treatment in mouse models of human colorectal cancer . A major drawback of peptides based ligands is their overall modest uptake. In this regard, small molecule caspase ligands offer an attractive alternative. The most prominent examples exist within the isatin sulfonamide moiety family.
Apoptosis Methods in Toxicology by Perpetua M. Muganda