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Western blot analysis of 14C-labelled membrane proteins.

A constant amount of liver microsomal proteins were incubated with different concentrations of a [14C]-inhibitor of a sugar transporter. Then, using a non-specific crosslinker, the protien-inhibitor complex was stabilised. The samples were loaded on to an 11% SDS-PAGE and blotted onto PVDF membrane. The Enhanced Autoradiography process invented by Frank Fox ( patent applied for) was then applied to the membrane.

western1.jpg (6925 bytes)The autoradiogram shown is obtained after 48 hours exposure of the membrane in a cassette at -800C. There is only one visible set of bands even after several weeks of exposure.

Identical results were previously obtained in our laboratory but after 3 - 4 weeks exposure with a BIO-RAD phosphor-imager or after 6 - 8 weeks exposure with conventional autoradiography film.

Dr. Ann Burchell. Department of Obstetrics and Gynaecology. Ninewells Hospital-Dundee University, Dundee, Scotland.

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