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.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004817:Epidermis / Q000032:analysis*; D000091344:Filaggrin Proteins; D006168:Guinea Pigs; D006801:Humans; D007381:Intermediate Filament Proteins / Q000032:analysis / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D011506:Proteins / Q000302:isolation & purification; D051381:Rats; D011919:Rats, Inbred Strains; D000099053:Histidine-Rich Glycoprotein
D000596:Amino Acids; C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D007381:Intermediate Filament Proteins; D011506:Proteins; D000099053:Histidine-Rich Glycoprotein