Summary: The NAD-dependent Aldehyde Dehydrogenase (ALDH) plays a vital role in cellular detoxification. It oxidizes various aldehydes and generates the corresponding carboxyolic acid. ALDH have been found in every cellular compartment. Based on its structure and function, ALDH comprises 3 major classes in mammals: Class 1 and Class 3 (the tumor form) are located in the cytosol and include both constitutive and induced forms; Class 2 is located in the mitochondria and only exists as the constitutive form. In humans, the ALDH superfamily consists of 19 genes. The mutation of ALDH genes (loss of function) causes human diseases such as Type II hyperprolinemia, pyridoxine-dependent seizure and hyperammonemia. Recent studies show that increased ALDH activity leads to several types of malignancies, serves as a cancer stem cell marker and correlates with poor prognosis. Therefore the early detection of ALDH activity levels can be prognostic and guide the therapeutic strategies. ALDH Activity Assay Kit