UID:
edoccha_9960073833802883
Format:
1 online resource (282 p.)
ISBN:
0-12-810118-0
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0-12-411623-X
Content:
Biochemical Ecotoxicology: Principles and Methods presents practical approaches to biochemical ecotoxicology experiments for environmental protection and conservation. With its methodical, stepped approach this essential reference introduces readers to current techniques for toxicity endpoint testing, suitable for laboratories of any size and budget. Each chapter presents a state-of-the-art principle, a quick and inexpensive procedure (including appropriate reagents), case studies, and demonstrations on how to analyze your results. Generic techniques are covered, suitable for a variety of
Note:
Description based upon print version of record.
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Front Cover; Biochemical Ecotoxicology; Copyright Page; Contents; List of Contributors; Dedication; Preface; Acknowledgments; Introduction; References; 1 Quantitative Assessments of Biochemical Analyses; 1.1 General Overview; 1.2 Replication in Data Analysis; 1.3 Replication of the Experiment; 1.3.1 Calibration or Standardization; 1.3.2 Using Specific and Generic Standards; 1.4 Reference Substances; 1.5 Defining Detection Limits in the Absence of Analytical Standards; 1.6 Reproducibility; 1.6.1 Reproducibility at the Calibration Level; 1.6.2 Reproducibility at the Level of the Experiment
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1.7 Using Reference Material to Standardize Methodologies1.8 Normalization Approaches; 1.8.1 Residual Extraction Method; 1.8.2 Stepwise Addition Method; References; 2 Tissue Preparation and Subcellular Fractionation Techniques; 2.1 Types of Homogenization Buffers; 2.1.1 pH; 2.1.2 Ion Chelators; 2.1.3 Antioxidants; 2.1.4 Proteases Inhibitors; 2.1.5 DNase/RNase Inhibitors; 2.1.6 Osmolarity; 2.1.7 Cryoprotective Agents; 2.2 Homogenization Methods; 2.2.1 Teflon Tissue Pestle; 2.2.2 Polytron; 2.2.3 Ultrasounds; 2.2.4 Freeze-Thaw; 2.2.5 Subcellular Fractionation by Differential Centrifugation
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2.2.6 Conservation of Biological Samples for BiomarkersReferences; 3 Preparation and Maintenance of Live Tissues and Primary Cultures for Toxicity Studies; 3.1 General Introduction; 3.2 Reagents Required and Solution Preparation; 3.3 Procedure; 3.3.1 Animal Collection and Maintenance; 3.3.2 Preparation of Primary Cell Cultures; 3.3.2.1 Dissection and Decontamination; 3.3.2.2 Tissue Dissociation; 3.3.2.3 Cell Suspension Culture; 3.3.2.4 Tissue Explant Culture; 3.4 Application to Toxicity Testing; 3.4.1 Cytotoxicity Evaluation; 3.4.1.1 Cell Viability Assessment
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3.4.2 Chronic Effects (Biomarker Expression)3.5 Calculations/Data Handling; 3.5.1 Calculating Cell Density and Viability Using the Neubauer Hemocytometer; References; 4 Measuring Effects at the Gene Transcription Level; 4.1 Introduction; 4.2 Material; 4.2.1 Reagents; 4.2.2 Equipment; 4.3 Procedure; 4.3.1 Choice of Detection Chemistry; 4.3.2 Primer Design; 4.3.3 Selecting the Target Nucleotide Sequence (Target and Reference Genes); 4.3.4 Preparation of Nucleic acid Template; 4.3.4.1 RNA Extraction from Cultured Cells; 4.3.4.2 Assessment of RNA Quality; 4.3.4.3 RNA Integrity Analysis
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4.3.4.4 RT4.3.5 Assay Optimization; 4.3.5.1 Primer Validation; 4.3.5.2 Standard Curve (Determination of Efficiency and Sensitivity); 4.3.5.3 Determination of Gene Transcripts by qPCR; 4.4 Data Analysis-Relative Quantification with Reference Gene Normalization; References; Web Resources; 5 Metal Metabolism and Detoxification; 5.1 Intracellular Free Metal Availability; 5.1.1 Introduction; 5.1.2 Reagents; 5.1.3 Procedure; 5.1.4 Data Calculation; 5.2 Subcellular Metal Binding Fractions; 5.2.1 Reagents and Equipment; 5.2.2 Procedure; 5.2.3 Data Analysis and Calculation; 5.3 Metallothioneins
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5.3.1 Silver Saturation Assay
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English
Additional Edition:
ISBN 0-12-411604-3
Additional Edition:
ISBN 1-306-94729-4
Language:
English
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