Stress proteins (HSP's): Methods of detection and their use as an environmental biomarker

This paper briefly reviews the nomenclature and application of stress prote ins as biomarkers, then gives a step by step description of the popular met hodologies used to measure stress protein in plant and animal tissues toget her with a critic of each approach and trouble shooting tips. Stress protei ns (previously termed "heat shock proteins") are induced by many environmen tal stresses including exposure to trace metals or organic pollutants, chan ges in temperature or osmolarity, hypoxia/anoxia, and exposure to ultraviol et radiation. Stress proteins, especially hsp70 and hsp60, have been used a s biomarkers in a range of algae, invertebrates, fish, and higher vertebrat es. Several suggestions are made to improve the utility of stress proteins as a biomarker of exposure e.g., consideration of the kinetics of stress pr otein induction relative to the pharmacokinetics of pollutant accumulation in the organism of concern, and selection of the type of stress protein for biomonitoring. The methodologies described include sample preparation, wes tern blotting, slot blotting, and antibody detection methods, especially fo r hsp70. Western blotting has been the most widely applied technique for as saying stress proteins but dot/slot blotting with the inclusion of an inter nal standard in each gel improves the approach by simplifying the technique -saving time and materials and producing a more quantitative result.