Conditions for reproducible detection of calmodulin and S100 beta in immunoblots.

Abstract:

: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 S100 beta 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.

authors

Van Eldik LJ,Wolchok SR

doi

10.1016/0006-291x(84)91022-2

subject

Has Abstract

pub_date

1984-11-14 00:00:00

pages

752-9

issue

3

eissn

0006-291X

issn

1090-2104

pii

0006-291X(84)91022-2

journal_volume

124

pub_type

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