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Focusing on essential insight into the methods and techniques required to dissect the complex mechanisms of NF- B activation, regulation, and function. NF-kappa B: Methods and Protocols guides readers through standard approaches to detect NF- B pathway activation, detection and analysis of NF- B signaling, and methods to study the control of NF- B signaling. As a volume in the highly successful Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and authoritative, NF-Kappa B: Methods and Protocols provides a timely and invaluable resource for researchers seeking to perform experiments aimed at understanding the role of NF- B signaling in health and disease.
List of contents
Electrophoretic Mobility Shift Assay Analysis of NF- B DNA Binding.- Detection of IkB Degradation Dynamics and IkB-a Ubiquitination.- Measurement of NF- B Transcriptional Activity and Identification of NF- B cis-regulatory Elements using Luciferase Assays.- Assessing Sites of NF- B DNA Binding using Chromatin Immunoprecipitation.- IKK Kinase Assay for Assessment of Canonical NF-kB Activation in Neurons.- Analysis of Epidermal Growth Factor-induced NF-kB Signaling.- Methods to Assess the Activation of the Alternative (non-canonical) NF-kB Pathway by Non-death TNF Receptors.- Systematic Detection of Non-canonical NF- B Activation.- Non-canonical NF- B activation and SDF-1 expression in Human Endothelial Cells.- Stable Reconstitution of IKK-deficient Mouse Embryonic Fibroblasts.- Dissecting NF- B Signaling Induced by Genotoxic Agents Via Genetic Complementation of NEMO-deficient 1.3E2 Cells.- Visualizing TCR induced POLKADOTS Formation and NF- B Activation in the D10 T Cell Clone and Mouse Primary Effector T cells.- Detection of Recombinant and Cellular MALT1 Paracaspase Activity.- TRAF Protein Function In Non-Canonical NF- B Signaling.- Roles of c-IAP Proteins in TNF Receptor Family Activation of NF-kB Signaling.- Elucidating Dynamic Protein-protein Interactions and Ubiquitination in NF-kB Signaling Pathways.- Immunoblot Analysis of Linear Polyubiquitination of NEMO.- In vitro Detection of NEMO-Ubiquitin Binding using DELFIA and Microscale Thermophoresis Assays.- Use of Fluorescence Spectroscopy For Quantitative Investigations Of Ubiquitin Interactions with the Ubiquitin-Binding Domains of NEMO.- Generation of a Proteolytic Signal: E3/E2-mediated Polyubiquitination of I B .- Control of NF- B Subunits by Ubiquitination.- Methodology to Study NF-kB/RelA Ubiquitination in vivo.- Using Sequential Immunoprecipitation and Mass Spectrometry to Identify Methylation of NF-kB.- Methods to Detect NF-kBAcetylation and Methylation.- A Method for the Quantitative Analysis of Stimulation-Induced Nuclear Translocation of the p65 subunit of NF-kB from Patient-Derived Dermal Fibroblasts.- Methods for Assessing the In Vitro Transforming Activity of NF- B Transcription Factor c-Rel and Related Proteins.- Using RNA Interference in Lung Cancer Cells to Target the IKK-NF- B Pathway.- Immunohistochemical Analysis of NF-kB in Human Tumor Tissue.- Assessment of Canonical NF-kB Activity in Canine Diffuse Large B-Cell Lymphoma.- NEMO Binding Domain Peptide Inhibition of Inflammatory Signal-Induced NF- B Activation In Vivo.- Regulation of NF B Signaling in Osteoclasts and Myeloid Progenitors.- NF- B Activation with Aging: Characterization and Therapeutic Inhibition.- The "Sneaking-ligand" Approach: Cell-type Specific Inhibition of the Classical NF-kB Pathway.- Sneaking-ligand Fusion Proteins Attenuate Serum Transfer Arthritis by Endothelium-targeted NF-kB inhibition.- Analysis of NF-kB Activation in Mouse Intestinal Epithelial Cells.- Characterizing the DNA Binding Site Specificity of NF-kB with Protein Binding Microarrays (PBMs).- Methods for Analyzing the Evolutionary Relationship of NF-kB Proteins Using Free, Web-driven Bioinformatics and Phylogenetic Tools.- Studying NFkB Signaling with Mathematical Models.
Summary
Focusing on essential insight into the methods and techniques required to dissect the complex mechanisms of NF-κB activation, regulation, and function. NF-kappa B: Methods and Protocols guides readers through standard approaches to detect NF-κB pathway activation, detection and analysis of NF-κB signaling, and methods to study the control of NF-κB signaling. As a volume in the highly successful Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and authoritative, NF-Kappa B: Methods and Protocols provides a timely and invaluable resource for researchers seeking to perform experiments aimed at understanding the role of NF-κB signaling in health and disease.
