Determination of Sialic Acids in Liver and Milk Samples of Wild-type and CMAH Knock-out Mice.
J Vis Exp
; (125)2017 07 14.
Article
en En
| MEDLINE
| ID: mdl-28745640
ABSTRACT
CMAH (cytidine monophosphate-N-acetylneuraminic acid hydroxylase) is responsible for the oxidation of cytidine monophosphate-N-acetylneuraminic acids in mammals. However, humans cannot oxidize cytidine monophosphate-N-acetylneuraminic acid to cytidine monophosphate-N-glycolylneuraminic acid due to a primary exon deletion of the CMAH gene. To understand the effects and implications of the lack of CMAH activity in more detail, a Cmah knock-out model in mice is of keen interest in basic and applied research. The analysis method to determine the phenotype of this mouse model is herein described in detail, and is based on the detection of both N-acetylneuraminic acid and N-glycolylenuraminic acid in the liver and milk of wild-type and Cmah knock-out mice. Endogenous sialic acids are released and derivatized with o-phenylenediamine to generate fluorogenic derivatives, which can be subsequently analyzed by HPLC. The presented protocol can be also applied for the analysis of milk and tissue samples from various other origins, and may be of use to investigate the nutritional and health effects of N-glycolylneuraminic acid.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Ácidos Siálicos
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Cromatografía Líquida de Alta Presión
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Leche
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Oxigenasas de Función Mixta
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Hígado
Límite:
Animals
Idioma:
En
Revista:
J Vis Exp
Año:
2017
Tipo del documento:
Article