Fumarase Activity Assay Kit (WST-8) 

Fumarase (Fumarate Hydratase, FH, EC 4.2.1.2) is an enzyme that catalyzes the reversible reaction of hydration and dehydration of fumarate to malate. It has two forms: mitochondrial and cytosolic forms. The mitochondrial form of fumarase is one of the key enzymes in citric acid cycle or tricarboxylic acid cycle (TCA), while cytosolic form is important for metabolism of amino acids and fumarate. In humans, fumarase deficiency leads to serious health problems such as fetal brain abnormality, hypotonia and renal cell carcinoma. Therefore, accurate measurement of fumarase activity is important for preventing, diagnosis and mechanistic study of fumarase deficiency.

MCE Fumarase Activity Assay Kit (WST-8) can be used to measure the amount of fumarase present in biological samples. The detection principle is as follows: Fumarase catalyzes the conversion of Fumarate into malate. The generated malate is then oxidized to oxaloacetic acid (OAA) in the presence of malate dehydrogenase (MDH). Simultaneously, NAD+ is reduced to NADH. The NADH interacts with the electron-coupled reagent (1-mPMS, 1-Methoxy-5-methylphenazinium Methyl Sulfate), transforms WST-8 into an orange-colored formazan displaying maximum absorption at 450 nm. In conclusion, there is a linear relationship between the amount of formazan generated in the system and the activity of fumarase in the sample.

Features of MCE Fumarase Activity Assay Kit (WST-8)

1. High Sensitivity: At 37˚C for 30 min, with the detection range of 0.7-30 U/L.

2. Wide Linear Range: At 37˚C for 30 min, the assay demonstrates a strong linear correlation within the concentration range of 20-1000 μM.

3. Fast Detection Speed and Broad Applicability: Suitable for various samples such as plasma, serum, red blood cells, tissues, and culture media.

-20℃, 1 year

Keep away from light and avoid repeated freeze-thaw cycles

NADH should be appropriately aliquoted and stored at -80°C after preparation into solution.