Retinoid Protocols (Methods in Molecular Biology Vol 89)

✍ Scribed by Redfern, Christopher

• Publisher: Humana Press
• Year: 1998
• Tongue: English
• Leaves: 423
• Edition: 1st
• Category: Library

✦ Synopsis

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.

✦ Table of Contents

Front......Page 1
1 Properties of Retinoids - Structure, Handling, and Preparation......Page 2
2 Quantitative Analyses of Naturally Occurring Retinoids......Page 28
3 Detection and Measurement of Retinoic Acid Production by Isolated Tissues Using Retinoic Acid-Sensitive Reporter Cell Lines......Page 40
4 lmmunohistochemistry for CRBPs and CRABPs......Page 54
5 Whole-Mount ln Situ Hybridization of Mouse Embryos Exposed to Teratogenic Levels of Retinoic Acid......Page 64
6 Reverse Transcriptase-Polymerase Chain Reaction (FIT-PCR) for Cellular Retinoid-Binding Proteins......Page 78
7 Methods for Producing Recombinant Human Cellular Retinaldehyde-Binding Protein......Page 88
8 Expression and Purification of CRABPs from E. coli......Page 102
9 Purification and Fluorescent Titration of Cellular Retinol-Binding Protein......Page 108
10 Fluorometric Titration of the CRABPs......Page 120
11 Expression and Mutagenesis of Retinol-Binding Protein......Page 138
12 Interactions of Retinol-Binding Protein with Transthyretin and Its Receptor......Page 152
13 Detection of Conformational Changes in Cellular Retinoid-Binding Proteins by Limited Proteolysis......Page 162
14 Measurement of Rates of Dissociation of Retinoids from the Interphotoreceptor Retinoid-Binding Protein......Page 174
15 Use of Antisense Oligonucleotides to Study the Role of CRABPs in Retinoic Acid-Induced Gene Expression......Page 188
16 Preparation of Polyclonal Antibodies to Retinoid Receptors......Page 200
17 Detection of RARs and RXRs in Cells and Tissues Using Specific Ligand-Binding Assays and Ligand-Binding lmmunoprecipitation Techniques......Page 214
18 Nonisotopic In Situ Hybridization for the Detection of Nuclear Retinoid Receptor Transcripts in Tissue Sections......Page 228
19 In Situ Hybridization with 35S-Labeled Probes for Retinoid Receptors......Page 242
20 Isolation of Retinoid Receptors from Mammalian Cells......Page 264
21 Analysis of Retinoid Receptor Phosphorylation......Page 272
22 Photoaffinity Labeling of RARs and Mapping of Labeled Sites by an Endoproteinase Combination Technique......Page 288
23 PCR Cloning of N-Terminal RAR lsoforms and APL-Associated PLZF-RARa Fusion Proteins......Page 302
24 RT-PCR in Diagnosis and Disease Monitoring of Acute Promyelocytic Leukemia (APL)......Page 328
25 A Two-Hybrid Protein Interaction System to Identify FactorsThat Interact with Retinoid and Vitamin D Receptors......Page 354
26 Gel-Shift Analysis and Identification of RXREs and RAREs by PCR-Based Selection......Page 372
27 Identification and Cloning of RA-Regulated Genes by mRNA-Differential Display......Page 384
28 Gene Targeting of Retinoid Receptors......Page 400