Immunostaining and Identification of Antigens
Tissue culture provides an opportunity to study the function of the nervous system under strictly controlled conditions. A major variable in neural tissue cultures, however, is the heterogeneity of the tissue used for the culture. Without knowing the relative proportions of neurons and glia, or the relative proportions of different subclasses of neurons, it can be difficult to interpret experimental results. Antibodies provide by far the most useful way of characterizing cells in culture. Using appropriate antibodies it is possible to determine the number of cells of a particular type in a culture, whether all of these cells have the same morphology or level of expression of antigen, and whether there is any preferential association between particular cell types. Because antibodies can be used in combination, it is possible to obtain an even more precise definition of cell type of developmental status. Although some of these measurements can be made using other methods, such as transmitter uptake, antibodies have two other important advantages. First, some antibodies can be used to compare cells in culture with those in intact tissue. Second, antibodies can frequently be used both to label cells in culture and to aid in biochemical isolation of the antigen.
KeywordsSodium Dodecyl Sulfate Wash Buffer Ammonium Persulfate Filter Strip Nitrocellulose Paper
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