Title:
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A study of the biochemistry and immunochemistry of differentiation of the normal epidermis and involved psoriatic epidermis
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The differentiation of skin during foetal
development, the maturation of cells in the normal adult
epidermis, and the alterations, occurring in psoriatic lesional
epiderms have been investigated using biochemical and
immunocytochemical methods. These were used as appropriate on
frozen sections, separated epidermis, dispersed keratinocytes and
subfractions of keratinocyte populations, prepared directly from
epidermis or epidermal cultures.
Various techniques were used to separate dispersed
keratinocytes, and these were monitored by light microscopy,
electron microscopy and immunocytochemistry. The method most
widely employed was Percoll density gradient centrifugation
(PDGC) . A potential Fluorescence activated cell sorter (FACS)
technique proved disappointing, but in a model system free flow
cell electrophoresis (FFCE) separated basal cell populations and
clearly has potential for use in man.
Analysis of protein components was performed using
2DGEL with a conventional rod and a new flatbed method for the
first dimension. various methods of protein detection were
investigated. Silver staining proved to be the most useful for
unlabelled proteins. Proteins labelled with 75Se or 35S
methionine were detected by autoradiography. Glycoproteins were
identified by lectin blotting of gels, by use of an enzyme
hydrazide technique or by incorporation of tritiated sugars and
subsequent fluorography or autoradiography. Hydrophobic proteins
were extracted using a Triton X-114 phase shift method following
by electrophoresis.
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