By Perpetua M. Muganda
This special quantity presents a unmarried, precious reference resource for ways that definitively establish and adequately quantify apoptosis. The publication starts off with universal tools applied to observe and quantitate apoptosis, in addition to apoptosis signaling pathways in toxicological and different comparable study. It maintains with multi-parametric and phased apoptosis assays for detecting early and past due apoptosis or distinguishing apoptosis from necrosis and autophagy. next chapters specialize in contemporary advances in actual time and high-throughput assays that become aware of and quantitate apoptosis and apoptosis signaling pathways. ultimate chapters specialize in contemporary advancements in preclinical anticancer therapeutics concentrating on apoptosis. Written for the Methods in Pharmacology and Toxicology sequence, chapters characteristic step by step descriptions of the methodologies, in addition to professional tips and implementation advice.
Vital and authoritative, Apoptosis equipment in Toxicology serves beginner scientists in addition to specialists, using more than a few tools from universal laboratory gear to high-end dear and automatic equipment able to appearing genuine time apoptotic measurements.
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Additional info for Apoptosis Methods in Toxicology
Bozza et al. pases measured in situ by binding of fluorochrome-labeled inhibitors of caspases (FLICA): correlation with DNA fragmentation. Exp Cell Res 259:308–313 Darzynkiewicz Z, Bedner E, Smolewski P, Lee BW, Johnson GL (2002) Detection of caspases activation in situ by fluorochromelabeled inhibitors of caspases (FLICA). Methods Mol Biol 203:289–299 Darzynkiewicz Z, Pozarowski P, Lee BW, Johnson GL (2011) Fluorochrome-labeled inhibitors of caspases: convenient in vitro and in vivo markers of apoptotic cells for cytometric analysis.
3 Detection of Apoptosis Control and toxicant exposed cells are assayed for apoptosis utilizing the three phased (early, intermediate, and late) apoptosis assays described below. The protocols outlined below utilize control and DEB-exposed cells assayed for apoptosis at various times post-DEB exposure. The Notes section (Notes 1–10) provides information for modifications needed for these assays to work in any system. 1 pSIVA-IANBD Assay, an Early Apoptosis Assay Cells are assayed for apoptosis utilizing the pSIVA-IANBD Apoptosis/Viability Microscopy Set Kit (Cat # IMG-6701 K, IMGENIX), as well as a modification of the kit manufacturer’s protocol, as described below.
Recent Pat Anticancer Drug Discov 4:28–35 7. Brunelle JK, Zhang B (2010) Apoptosis assays for quantifying the bioactivity of anticancer drug products. Drug Resist Updat 13:172–179 8. Kroemer G, Galluzzi L, Vandenabeele P, Abrams J, Alnemri ES, Baehrecke EH et al (2009) Classification of cell death: recommendations of the Nomenclature Committee on Cell Death 2009. Cell Death Differ 16:3–11 9. Eum KH, Lee M (2011) Crosstalk between autophagy and apoptosis in the regulation of paclitaxel-induced cell death in v-Ha-rastransformed fibroblasts.