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This volume provides detailed protocols for the performance, analysis, and troubleshooting of in vitro and in vivo experiments related to programmed cell death. Chapters compile conventional techniques such as western blot and qPCR and state-of-the art transmission electron microscopy and real-time multiplexed imaging assays. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and accessible, Detection of Cell Death Mechanisms: Methods and Protocols aims to ensure seamless execution of protocols on specific cell death type.
1. Determination of Caspase Activation by Western Blot
Hussein Chehade, Alexandra Fox, Gil Mor, and Ayesha B. Alvero
2. Detection of Unfolded Protein Response by Polymerase Chain Reaction
Alexander Paridon, Alexandra Fox, and Ayesha B. Alvero
3. Subcellular Fractionation to Demonstrate Activation of Intrinsic Apoptotic Pathway
Hussein Chehade, Alexandra Fox, Gil Mor, and Ayesha B. Alvero
4. A Triple-Parameter Based Laboratory Friendly Fluorescence Imaging to Identify Apoptosis in Live Cells
Pradip De, Jennifer Carlson Aske, and Nandini Dey
5. Flow Cytometric Analyses of p53-mediated Cell Cycle Arrest and Apoptosis in Cancer Cells
Nour N. Al Zouabi, Cai M. Roberts, Z. Ping Lin, and Elena S. Ratner
6. A Real-Time, Bioluminescent Apoptosis Assay
Kevin R. Kupcho and Andrew L. Niles
7. Detection of Anoikis Using Cell Viability Dye and Quantitation of Caspase Activity
Roslyn Tedja, Alexandra Fox, and Ayesha B. Alvero
8. Time and Dose Dependent Toxicity Studies in 3D Cultures Using a Luminescent Lactate Dehydrogenase Assay
Natasha Karassina, Peter Hofsteen, James Cali, and Jolanta Vidugiriene
9. Visualization and Quantification of Neutrophil Extracellular Traps
Mancy Tong and Vikki M Abrahams
10. 2In Vitro Identification and Isolation of Human Neutrophil Extracellular Traps
Guillermina Calo, Analia Silvina Trevani, Esteban Grasso, Irene Angelica Keitelman, Rosanna Ramhorst, Claudia Pérez Leirós, and Florencia Sabbione
11. Induction and Detection of Necroptotic Cell Death in Mammalian Cell Culture
Mikhail Chesnokov, Imran Khan, and Ilana Chefetz
12. Visualization of Necroptotic Cell Death through Transmission Electron Microscopy
Naresh Golla, Linda J. Hong, and Ilana Chefetz
13. Methodology for Comprehensive Detection of Pyroptosis
Yang Feng and Xiaoli Huang
14. Determination and Quantitation of Cytotoxic T Cell-Mediated Cell Death
Han-Hsuan Fu and Harry Qui
15. Detection of Immunogenic Cell Death in Tumor Vaccination Mouse Model
Kazuki Tatsuno, Patrick Han, Richard Edelson, and Douglas Hanlon
16. A Luminescent Assay to Quantify Cell Viability in Real Time
Peter Hofsteen, Natasha Karassina, James J. Cali, and Jolanta Vidugiriene
17. Kinetic Measurement of Apoptosis and Immune Cell Killing Using Live-Cell Imaging and Analysis
Jill E. Granger and Daniel M. Appledorn
18. Two Plasmid-Based Systems for CRISPR/Cas9 Mediated Knockout of Target Genes
Cai M Roberts and Elena S Ratner
19. Determining Cell Death Pathway and Regulation by Enrichment Analysis
Katherine Gurdziel
20. Transcription Factor Binding Site Identification and Enrichment Analysis
Joe L. Guy and Gil Mor
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