Poly(ADP-Ribose) polymerase : methods and protocols / edited by Alexei V. Tulin. -- Third edition. -- New York, NY : Humana Press, 2023. – (58.17/M592/v.2609)

Contents

Preface
Contributors
PART I  DETECTION AND QUANTIFICATION OF PADPR
1  Fluorescence-Based Analyses of Poly(ADP-Ribose) Length by Gel Electrophoresis, High-Performance Liquid Chromatography, and Capillary Electrophoresis
2  Detecting and Quantifying pADPr In Vivo
3  Quantitative Analysis of Nuclear Poly(ADP-Ribose) Dynamics in Response to Laser-Induced DNA Damage
4  Analyzing PARP1 Activity: Small Molecule Reactants and Attached Chains of Poly (ADP-Ribose)
5  Detecting Poly (ADP-Ribose) In Vitro and in Cells Using PAR Trackers
6  An Enzyme-Linked Immunosorbent Assay to Quantify Poly (ADP-Ribose) Level In Vivo
7  Subcellular Quantitation of ADP-Ribosylation by High-Content Microscopy
8  A Simple Method to Study ADP-Ribosylation Reversal: From Function to Drug Discovery
PART II  IDENTIFICATION OF PROTEIN TARGETS
9  Immunoprecipitation Using Mono-ADP-Ribosylation-Specific Antibodies
10  A Clickable NAD+ Analog-Based Assay of Poly(ADP-Ribosyl)ated Proteins
11  Functional Analysis of Histone ADP-Ribosylation In Vitro and in Cells
PART III  FUNCTIONAL ANALYSIS
12  Studying the Immunomodulatory Functions of PARP1 and PARP2 in Mouse Models of Cancer
13  Methods for Investigating Transient Receptor Potential Melastatin-2 (TRPM2): A Cation Channel Activated by ADP-Ribose and Involved in Cell Death
14  Methods to Assess the Role of PARPs in Regulating Mitochondrial  Oxidative Function
15  Characterizing ADP-Ribosylation Sites Using Af1521 Enrichment Coupled to ETD-Based Mass Spectrometry
16  Cytological Approaches to Visualize Intracellular Dynamics of  RNA-Binding Proteins at Active Genes in Drosophila
PART IV  POLY(ADP-RIBOSYL)ATING PATHWAY IN CHROMATIN AND GENE EXPRESSION
17  Chromatin Immunoprecipitation Approach to Determine How PARP1 Domains Affect Binding Pattern to Chromatin
18  Approach to Measuring the Effect of PARP1 on RNA Polymerase II Elongation Rates
19  Examining the Effect of PARP-1 Inhibitors on Transcriptional Activity of Androgen Receptor in Prostate Cancer Cells
PART V  USING ANIMAL MODELS
20  Using Drosophila Genetics to Identify Factors that Affect PARP1 Activity In Vivo
21  Generating PARP Knockout D. melanogaster with CRISPR/Cas9 System
22  TaqMan Multiplex qPCR Method to Genotype PARG Knockout Mice
PART VI  DESIGNING AND TESTING PARP-1 INHIBITORS
23  Cell-Based Screening for New PARP Inhibitors Utilizing PARG-Mutated Mouse Embryonic Stem Cells
24  Quantification of PARP7 Protein Levels and PARP7 Inhibitor Target Engagement in Cells Using a Split Nanoluciferase System
PART VII  TARGETING THE UNDERSTUDIED COMPONENTS OF PADPR PATHWAY
25  Purification of Recombinant Human PARG and Activity Assays
26  Purification of Recombinant Human PARP-3
Index