Golgi detection in mitotic and interphase cells by antibodies to secreted galactosyltransferase.
Secreted galactosyltransferase from bovine milk was used to induce antibodies cross-reacting with corresponding intracellular enzymes in a variety of cell lines and tissues. In contrast to the original antigen, the reactive intracellular galactosyltransferase appears as individual species (apparent MW approx. 42000-46000) in SDS-polyacrylamide gel electrophoresis. In indirect immunofluorescence microscopy affinity-purified IgGs locate the galactosyltransferase in a distinct perinuclear and juxtanuclear position indicative for the Golgi region. The rearrangement of labelled structures upon colcemid or monensin treatment--drugs known to influence Golgi morphology and function--is further proof for a Golgi association. The fate and distribution of Golgi elements during mitosis is described at the light microscopical level using galactosyltransferase as easily identifiable marker. In addition we evaluate the utilization of wheat germ agglutinin (WGA) binding for Golgi identification on tissue culture cells and show that WGA is not a reliable marker for certain cell types such as MDCK.
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D005700:Galactosyltransferases / Q000276:immunology*; D006056:Golgi Apparatus / Q000648:ultrastructure*; D006801:Humans; D007106:Immune Sera; D007399:Interphase; D008938:Mitosis
D007106:Immune Sera; D005700:Galactosyltransferases