Conditions for reproducible detection of calmodulin and S100β in immunoblots

Linda J. Van Eldik, Sandra R. Wolchok

Research output: Contribution to journalArticlepeer-review

100 Scopus citations


The conditions used in some immunoblot procedures can fail to detect calmodulin, S100 proteins, and other proteins with similar physical properties. We describe here some of the basis of this difficulty, and provide an immunoblot protocol that allows the rapid and reproducible detection of calmodulin and S100β in crude biological samples. These proteins are rapidly transferred from sodium dodecyl sulfate-polyacrylamide gels to membrane matrices, and retention on the matrix is enhanced by a glutaraldehyde fixation step. Either nitrocellulose or a positively charged membrane filter (ZetaProbe) can be used as the immobilizing matrix. By combining microslab gel electrophoresis, 30 min electrophoretic transfer, and glutaraldehyde fixation of nitrocellulose paper, an immunoblot analysis can be done in an 8-hr day.

Original languageEnglish
Pages (from-to)752-759
Number of pages8
JournalBiochemical and Biophysical Research Communications
Issue number3
StatePublished - Nov 14 1984

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology


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