Comparison of different isolation methods for catalase activity measurement in adipose tissue

Recent studies have shown that adipose tissue is not only an energy store, but also an endocrine organ. White adipose tissue, which is abundant in the body, stores excess energy in triglyceride form and contributes to energy homeostasis. Obesity caused by the increased adipose tissue mass related with oxidant antioxidant in balance and low grade chronic inflammation. Antioxidant enzymes are proteins that help suppress the oxidative stress by causing less active radical formation or by reducing the damage of the free radical chain reaction on proteins, lipids, carbohydrates and DNA. The catalase enzyme breaks down H2O2, an important source of oxidant, into water and oxygen, thus preventing the formation of oxidative stress. As fat tissue has high lipid and low protein content which leads to high lipid interference. Hence, protein isolation and measurement are difficult in fat tissue. In this study, we aimed to investigate the effect of using different protein isolation methods from adipose tissue on eventual CAT activity and to determine the necessary conditions for measurement. . Catalase activity was measured by Aebi method. Retroperitoneal fat tissue was obtained from the rats and used for three different homogenization methods. In homogenization 1 (H1) chloroform / methanol was used as the organic solvent, in homogenization 2 (H2) and in homogenization 3 (H3) only chloroform was used. The result of the evaluations revealed that the average specific activity values of H1 method were elevated than the other two methods. It was concluded that H1 method in retroperitoneal adipose tissue may be more suitable for CAT enzyme activity measurement.