The specific method use to determine N-acetylneuaminic acid (NANA) is considered proprietary. However, the assay is a modification of a Neuraminidase enzymatic assay. A prepared fetuin solution is hydrolyzed chemically to determine the "total NANA".
Two concentrations of a NANA standard are utilized, 0.150 mg/ml for total NANA and 0.06 mg/ml for free NANA, and test is monitored at 550 nm.
The free NANA, monitored at 340 nm, is done to confirm results.
Please see the link below to review additional information available in the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/127/380/f2379pis.pdf



