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1. Neither cold acclimation (2–3°C) nor laboratory captivity affected the amount of hsp 70 in the lungless salamander Plethodon cinereus. 2. Plethodon jordani had substantial amounts of heat shock protein (hsp) 70 present in its tissues when acclimated to 15°C (LD 12:12). 3. The increase in hsp 70 in P. jordani (13.2%) after heat shock at 2°C below its critical thermal maximum (CTMax) (34.1°C) was not statistically significantly higher than the control value. 4. An immunoblot with monoclonal antibody after 2-D SDS PAGE gel indicated that constitutive hsp 70 was the major form of hsp 70 in P. jordani.

1. 1.|Fish (Pimephales promelas, Salmo trutta, Ictalurus natalis and Ambloplites rupestris) respond to seasonal changes in their environment by synthesizing heat shock proteins with a molecular weight of approximately 70 Kd (hsc/hsp-70). 2. 2.|Lowest levels of hsc/hsp-70 measured with ELISA occurred in winter, followed by a high level in spring, a significant decrease in summer and another in fall. 3. 3.|Caution must be applied in developing heat shock proteins as biomarkers so that their presence in organisms under “normal” conditions is not misinterpreted as an indication that the organisms are stressed.

1. 1. We purified hsp-70 from salamander, Plethodon cinereus, and established mouse myeloma-B lymphocyte hybridoma cell lines to produce monoclonal antibodies against hsp-70. 2. 2. An ELISA test using the monoclonal antibody allows rapid quantification of hsp-70 from tissue homogenates from large numbers of animals. 3. 3. Salamanders (P. cinereus, Eurycea bislineata, Desmognathus ochrophaeus, Ambystoma talpoidium), turtle (Trachemys scripta) and fish (Fundulus heteroclitus, Pimephales promelas) contain large amounts of constitutive hsp-70 and a small but consistent increase in hsp-70 within 1 h of heat shock. 4. 4. ELISA test indicates that there is considerable variation in hsp-70 level among individuals of a species. 5. 5. Constitutive hsp-70 is present at normal ecological temperatures and additional hsp-70 is induced at temperatures as low as 25°C in A. talpoideum.