engleski

Ganglioside immunohistochemistry - shooting at a moving target

Numerous studies of ganglioside distribution have been published, but their results were somewhat conflicting and the situation is still not clear. Generally the inconsistencies in observed distribution were considered to be a consequence of different methods of fixation and varying specificity of antibodies, but in our hands even the same fixation procedure and highly specific monoclonal IgG antibodies were still producing inconsistent results. After numerous experiments we have realized that ganglioside immunohistochemistry is particularly sensitive to various influences. Even smallest amounts of organic solvents or detergents in any stage of the procedure (including fixation in cold solvents) will extract significant amounts of gangliosides from cell membranes and also enable their transfer from one cell, or tissue to another. For reproducible results all steps of ganglioside immunohistochemistry have to be performed at +4º C, because at higher temperatures gangliosides shed significantly. Even when all steps are performed in cold and with utmost care, ganglioside immunohistochemistry still resembles shooting at moving targets, because gangliosides also jump from one cell or tissue to another in most commercially available mounting media. In some of them this happens in a matter of hours, in some in a matter of days, but the picture is constantly changing. To make things even more complicated, different gangliosides jump to and from different structures, and some jumping can also be observed for GPI-linked proteins.

Gangliosides; immunohistochemistry

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