Immunostaining and Identification of Antigens

  • Colin J. Barnstable


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.


Sodium Dodecyl Sulfate Wash Buffer Ammonium Persulfate Filter Strip Nitrocellulose Paper 
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Further Reading

  1. Akagawa, K. and Barnstable, C. J. (1986), Identification and characterization of cell types in monolayer cultures of rat retina using monoclonal antibodies. Brain Res. 383, 110–120.PubMedCrossRefGoogle Scholar
  2. Andrews, A. T. (1986), Electrophoresis. Theory, Techniques, and Biochemical and Clinical Applications. 2nd ed., Oxford University Press.Google Scholar
  3. Arimatsu, U., Naegele, J. R., and Barnstable, C. J. (1987), Molecular markers of neuronal subpopulations in layers, 4, 5, and 6 of cat primary visual cortex. J. Neurosci. 7, 1250–1263.PubMedGoogle Scholar
  4. Barnstable, C. J. (1985), Monoclonal antibodies as molecular probes of the nervous system, in Hybridoma Technology in the Biosciences and Medicine, Springer, T., ed., Plenum, New York, pp. 269–289.CrossRefGoogle Scholar
  5. Barnstable, C. J. (1987), Immunological studies of the diversity and development of the mammalian visual system. Immunol. Reviews 100, 47–78.CrossRefGoogle Scholar
  6. Childs, G. V. (1986), Immunocytochemical Technology. Liss, New York.Google Scholar
  7. Constantine-Paton, M., Blum, A. S., Mendez-Otero, R., and Barnstable, C. J. (1986), A cell surface molecule distributed in a dorsoventral gradient in the perinatal rat retina. Nature 324, 459–462.PubMedCrossRefGoogle Scholar
  8. Dulbecco, R. and Vogt, M. (1954), Plaque formation and isolation of pure lines with poliomyelitis virus. J. Exp. Med. 99, 167–182.PubMedCrossRefGoogle Scholar
  9. Harlow, E. and Lane, D. (1988), Antibodies. A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
  10. Laemmli, U. K. (1970), Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.PubMedCrossRefGoogle Scholar
  11. Towbin, H., Staehelin, T., and Gordon, J. (1979) Electrophoretic transfer of proteins from Polyacrylamide gels to nitrocellulose sheets. Proc. Natl. Acad. Sci. USA 76, 4350–4354.PubMedCrossRefGoogle Scholar
  12. Zaremba, S., Naegele, J. R., Barnstable, C. J., and Hockfield, S. (1990), Neuronal subsets express multiple high-molecular-weight cell-surface glycoconjugates defined by monoclonal antibodies Cat-301 and VC 1.1. J. Neurosci. 10, 2985–2995.PubMedGoogle Scholar

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© Springer Science+Business Media New York 1997

Authors and Affiliations

  • Colin J. Barnstable

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