Peroxisomes

I: Isolation and Characterization of Peroxisomes

- General Indroduction to Isolation and Characterization of Peroxisomes

- Experiment 1. Preparation and Purification of Peroxisomes; Subfractionation of Purified Peroxisomes; Acyl-CoA Oxidase Activity Measurements

- Experiment 2. Immunoblotting of Peroxisomal Proteins with Monospecific Antibodies

- Experiment 3. Morphology of Peroxisomes in Light- and Electron Microscopy

II: Molecular Biology

- Experiment 4. Northern Blotting Analysis of Rat Liver mRNA Encoding for Peroxisomal Acyl-CoA Oxidase

- Experiment 5. Transient Co-transfection Assay: Measure of the Chloramphenicol Acetyl Transferase (CAT) Activity in Cytosol Extracts from Transfected Cells

- Experiment 6. Synthesis of Oligonucleotides Used as Probes; Purification by HPLC

- Experiment 7. Computer Analysis of DNA and Protein Sequences

III: Toxicology and Pharmacology

- Experiment 8. Cytochrome P450 Isoenzyme Induction Pattern in Rats Treated with Peroxisome Proliferators and Classical Inducers

- Experiment 9. Measurement of the in Vitro Glucuronidation of Peroxisome Proliferator Carboxylic Acids by Liver Microsomes and by Genetically Modified V 79 Cell Line

- Experiment 10. Molecular Modeling and Drug Design: Application to Some Peroxisome Proliferator Agents

IV: Cell Culture and Genetic Diseases

- Experiment 11. Fao Cell Line as a Model for the Study of the Effect of Peroxisome Proliferators on Cellular Functions

- Experiment 12. Measurement of Dihydroxyacetone-Phosphate Acyl-Transferase (DHAP-AT) Activity in Liver Peroxisomes and Cell Lines

- Experiment 13. In Vitro Transformation of Human Fibroblasts with SV40 T Antigen (Lipofection)

- Experiment 14. Somatic Cell Hybridization as a Tool for Genetic Analysis of Peroxisomal Activities

- Supplement

Peroxisomes - or microbodies - are organelles present in nearly all cells. Interest stems not only from their various biological and cell physiological roles but also because several genetic diseases are known with deficient peroxisomal functions. In response to several drugs or pesticides, peroxisomes start to proliferate. Due to this property they may also be used as toxicological indicators. Designed for use at the lab bench, this manual is ideal for everyone involved in peroxisome research or concerned with hepatotoxicity testing of new compounds.