A comprehensive collection of state-of-the-art biochemical and molecular methods for retinoid research. These easily reproducible techniques range from methods of handling and analyzing retinoids to advanced protocols for gene targeting, from binding protein function in transgenic animals to the study of nuclear retinoid receptors. In addition, the book makes accessible to all retinoid researchers such advanced techniques as cloning via RT-PCR, fluorimetry methods, recombinant protein purification and characterization, the use of antisense oligonucleotides to study the role of CRABPs, immunological and in situ hybridization methods, photoaffinity labeling, gel-shift analysis, and differential display.
Part I. Handling and Analysis of Retinoids. Properties of Retinoids:Structure Handling and Preparation Arun B. Barua and Harold C. Furr. Quantitative Analyses of Naturally Occurring RetinoidsJoseph L. Napoli and Ronald L. Horst. Detection and Measurement of Retinoic Acid Production by Isolated Tissues Using Retinoic Acid-Sensitive Reporter Cell Lines Michael Wagner. Part II. Retinoid-Binding Proteins. Immunohistochemistry for CRBPs and CRABPsUlf Eriksson and Anne-Lee Gustafsson. Whole-Mount In Situ Hybridization of Mouse Embryos Exposed to Teratogenic Levels of Retinoic AcidSally D. Lyn. Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) for Cellular Retinoid-Binding Proteins Andrew D. Loughney and Christopher P. F. Redfern. Methods for Producing Recombinant Human Cellular Retinaldehyde-Binding Protein John W. Crabb Yang Chen Steve Goldflam Karen West and James Kapron. Expression and Purification of CRABPs from E. coli Joseph L. Napoli Zuzana Sperkova Marek Sperka and Paul D. Fiorella. Purification and Fluorescent Titration of Cellular Retinol-Binding Protein Giorgio Malpeli Claudia Folli Davide Cavazzini Giovanni Sartori and Rodolfo Berni. Fluorometric Titration of the CRABPs Andrew W. Norris and Ellen Li. Expression and Mutagenesis of Retinol-Binding Protein Manickavasagam Sundaram Asipu Sivaprasadarao and John B. C. Findlay. Interactions of Retinol- Binding Protein with Transthyretin and Its Receptor Asipu Sivaprasadarao Manickavasagam Sundaram and John B. C. Findlay. Detection of Conformational Changes in Cellular Retinoid- Binding Proteins by Limited Proteolysis Robert S. Jamison Marcia E. Newcomer and David E. Ong. Measurement of Rates of Dissociation of Retinoids from the Interphotoreceptor Retinoid-Binding Protein Noa Noy. Use of Antisense Oligonucleotides to Study the Role of CRABPs in Retinoic Acid- Induced Gene Expression Paul Nugent and Robert M. Greene. Part III. Nuclear Retinoid Receptors. Preparation of Polyclonal Antibodies to Retinoid Receptors Jacqueline A. Dyck and Vickie J. LaMorte. Detection of RARs and RXRs in Cells and Tissues Using Specific Ligand-Binding Assays and Ligand-Binding Immunoprecipitation Techniques Elizabeth A. Allegretto. Nonisotopic In Situ Hybridization for the Detection of Nuclear Retinoid Receptor Transcripts in Tissue Sections Xiaochun Xu and Reuben Lotan. In Situ Hybridization with S-Labeled Probes for Retinoid Receptors Karen Niederreither and Pascal Doll'. Isolation of Retinoid Receptors from Mammalian Cells Ann K. Daly and Christopher P. F. Redfern. Analysis of Retinoid Receptor Phosphorylation Ali Tahayato Christophe Rachez Pierre Formstecher and Philippe Lefebvre. Photoaffinity Labeling of RARs and Mapping of Labeled Sites by an Endoproteinase Combination TechniqueYuichi Hashimoto and Toru Sasaki. PCR Cloning of N-Terminal RAR Isoforms and APL-Associated PLZF-RARa Fusion Proteins Arthur Zelent. RT-PCR in Diagnosis and Disease Monitoring of Acute Promyelocytic Leukemia (APL) David Grimwade Stephen Langabeer Kathy Howe and Ellen Solomon. A Two- Hybrid Protein Interaction System to Identify Factors That Interact with Retinoid and Vitamin D Receptors Paul N. MacDonald. Gel-Shift Analysis and Identification of RXREs and RAREs by PCR- Based Selection Myriam I. Baes and Peter E. Declercq. Identification and Cloning of RA-Regulated Genes by mRNA Differential Display Jay A. White and Martin Petkovich. Gene Targeting of Retinoid Receptors David Lohnes. Index.
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