Oct 1, 1987

Human hematoxylin-stainable protein of keratohyalin granules origin. I. Extraction and purification

The Journal of Investigative Dermatology
T Tezuka, M Takahashi

Abstract

A human hematoxylin-stainable protein (HSP) was extracted from the massive stratum corneum of epidermal cysts. This protein was purified in two steps; first, through preparative isoelectric focusing, and, second, by affinity column chromatography bound with the specific, monoclonal antibody to keratohyalin granules (Ted-H-1). In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the purified HSP consisted of two proteins (70 and 62 kilodaltons [kd]), but only the 62-kd protein was detected in the 15,000 g supernatant fraction of the extract using the immunoblotting technique. The amino acid composition of the purified HSP included 30% glycine, 15% serine, 12% glutamic acid, and 4% ornithine, but only 2.3% histidine. Using the indirect immunofluorescent technique, observation showed that the monoclonal antibody, Ted-H-1, to the HSP, formed as a result of the partially purified antigen, was located in three places: 1) the keratohyalin granules; 2) in the cell membrane region of the lower part of the stratum corneum of the skin samples (forearm, cheek, and back); and 3) the keratohyalin granules in the follicular epithelium and on the trichohyalin granules. Reaction product was not seen in either the acrosyringium or in th...Continue Reading

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Mentioned in this Paper

Monoclonal Antibodies
Immunofluorescence Assay
Histidine
SDS-PAGE
Stratum Granulosum
Tromethamine
Epithelium
Cheek Structure
Glycine
Immunoblotting

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