A microvolume protocol for catalase activity

We developed and validated a microvolume protocol for measuring catalase (CAT) activity using a NanoDrop spectrophotometer, enabling reliable quantification with as little as 2 µL of sample. This methodology provides an efficient alternative to conventional assays that require substantially larger volumes and is particularly suited for studies with limited biological material, such as small organisms (Panagrellus redivivus and Caenorhabditis elegans) or specific organs of Drosophila. Our results demonstrate that the protocol accurately evaluates CAT activity, reflecting the physiological and metabolic characteristics of the analyzed tissues. To ensure specificity, CAT inhibitors and protein denaturation controls were employed. Overall, this methodology expands experimental possibilities for assessing enzymatic activity in reduced biological models and investigating their responses to oxidative stress.