Organelle-specific antibodies: production of antibodies to Golgi subcompartments

We have devised a strategy for producing Golgi subcompartment-specific antibodies that involves utilizing Triton X-114-phase separated membrane proteins derived from rat pancreatic Golgi subfractions as immunogens. When we tested the approach by immunizing rabbits with membrane proteins derived from...

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Bibliographic Details
Published inProgress in clinical and biological research Vol. 270; p. 129
Main Authors Saraste, J, Bronson, M, Palade, G E, Farquhar, M G
Format Journal Article
LanguageEnglish
Published United States 1988
Subjects
Animals
Antibodies - immunology
Cell Fractionation
Golgi Apparatus - immunology
Golgi Apparatus - ultrastructure
Pancreas - ultrastructure
Rats
Subcellular Fractions - ultrastructure
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Summary:We have devised a strategy for producing Golgi subcompartment-specific antibodies that involves utilizing Triton X-114-phase separated membrane proteins derived from rat pancreatic Golgi subfractions as immunogens. When we tested the approach by immunizing rabbits with membrane proteins derived from heavy Golgi subfractions that are known to be enriched in cis Golgi elements, we succeeded in generating an antibody that recognized a 58 kD protein that was restricted in its distribution to cis Golgi cisternae in several cell types. Thus we have demonstrated the feasibility of the approach we devised for generation of Golgi subcompartment-specific antibodies, and we have also succeeded in identifying a heretofore unknown cis Golgi marker protein.
ISSN:0361-7742