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Storage stability of hepatobiliary enzymes such as, Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Alkaline phosphatase (ALP) and Gamma glutamyltransferase (GGT) in the sera of dogs of mixed breeds reared under the hot humid climatic conditions of Kerala was assessed. All the animals were apparently healthy, free of external parasites, dewormed regularly and vaccinated routinely against infectious diseases. Concentrations of ALT, AST, ALP and GGT were determined using an automated blood analyzer. Stabilities of enzymes in sera stored at room temperature (22 to 27°C), 4°C and -20°C were compared with the enzyme activities of fresh sera (day 0) on day 1, 2, 5, 8, 11 and 14. Alanine aminotransferase was found to be sufficiently stable up to 14 days at 4 °C, whereas it was stable only for one day at -20°C and was totally unstable at room temperature. Aspartate aminotransferase was stable both at 4 °C and -20°C up to 14 days whereas, it was totally unstable at room temperature. Alkaline phosphatase showed great variation upon storage when compared to other hepatobiliary enzymes and it is suggested that its estimation should be performed in fresh sera to get an accurate result even (hough the study suggested -20°C as better. Gamma glutamyltransferase was found to be highly stable at room temperature, 4°C and -20°C during the entire experimental period. It is therefore advisable to consider stability of each serum hepatobiliary enzyme for different animals separately before preserving sera to get more valid and reliable results.