A high performance liquid chromatography method to obtain rat epidermal filaggrin

Sodium pyrophosphate-glycerol buffer, a non-denaturing buffer has been used to solubilize epidermal proteins. The extracted proteins are different in their electrophoretic profile in various mammalian species, 47 K in rat and guinea pig, 31 K and 60 K in mouse and 43 K and 60 K proteins in human epidermis were most predominant. Electrophoretic analyses show synthesis of new proteins as a function of age in rat tissue. Purification of the major rat protein fraction was achieved using a reverse phase high-performance liquid chromatography using a gradient of 5-50% acetonitrile. Based upon the molecular size, amino acid data and immunodiffusion analysis, we conclude the purified rat protein to be filaggrin.