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. 1982 Nov;16(5):803–807. doi: 10.1128/jcm.16.5.803-807.1982

Application of potassium thiocyanate as a destaining agent for confirming the specificity of viral immunofluorescence.

L W Greenham, E O Caul
PMCID: PMC272480  PMID: 6185529

Abstract

A study was made of the destaining effects of various agents on measles-infected monolayers which were stained by the indirect method of immunofluorescence. It was found that KSCN, a chaotropic agent, gave the required destaining but was nonspecific in its action, removing antigen as well as adsorbed immunoglobulins. By using a KSCN-EDTA destaining solution, it was shown that two sequential restainings of infected monolayers were possible, which permitted the demonstration of the specificity of the observed immunofluorescence. The technique may be of particular use with clinical specimens in which relatively few cells give immunofluorescence and when the specificity of this fluorescence is questioned.

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Selected References

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  1. Dandliker W. B., Alonso R., de Saussure V. A., Kierszenbaum F., Levison S. A., Schapiro H. C. The effect of chaotropic ions on the dissociation of antigen-antibody complexes. Biochemistry. 1967 May;6(5):1460–1467. doi: 10.1021/bi00857a031. [DOI] [PubMed] [Google Scholar]
  2. Fraser K. B., Armstrong M., Shirodaria P. V., Gharpure M. Affinity for measles virus anti-haemolysin of a residual immunoglobulin M in sera of some patients with multiple sclerosis. Clin Exp Immunol. 1979 May;36(2):304–310. [PMC free article] [PubMed] [Google Scholar]
  3. Hatefi Y., Hanstein W. G. Solubilization of particulate proteins and nonelectrolytes by chaotropic agents. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1129–1136. doi: 10.1073/pnas.62.4.1129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Heimer G. V., Taylor C. E. Improved mountant for immunofluorescence preparations. J Clin Pathol. 1974 Mar;27(3):254–256. doi: 10.1136/jcp.27.3.254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. KLEINSCHMIDT W. J., BOYER P. D. Interaction of protein antigens and antibodies. I. Inhibition studies with the egg albumin-antiegg albumin system. J Immunol. 1952 Sep;69(3):247–255. [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
  8. Ploem J. S. The use of a vertical illuminator with interchangeable dichroic mirrors for fluorescence microscopy with incidental light. Z Wiss Mikrosk. 1967 Nov;68(3):129–142. [PubMed] [Google Scholar]
  9. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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