Immunoblotting conditions for human hemoglobin chains

Yusuke Suzuki, Yoshihiko Takeda, Tohru Ikuta

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Immunoblotting to analyze low-molecular-weight proteins like calmodulin and metallothioneins is challenging and requires modifications for reproducible detection. Human globin chains are 17-kDa proteins and are not detectable by conventional immunoblotting using nitrocellulose membranes. Here we describe an immunoblotting method using nitrocellulose membranes that allows quantitative analyses of globin chains. Although previous studies have demonstrated that the fixation of blotted membranes with glutaraldehyde improves immunodetection of low-molecular-weight proteins, we found that the detection sensitivity for human globins is increased markedly by fixation with paraformaldehyde, but not glutaraldehyde. This immunoblotting procedure facilitates studies of posttranscriptional mechanisms for globin gene expression.

Original languageEnglish (US)
Pages (from-to)218-220
Number of pages3
JournalAnalytical Biochemistry
Volume378
Issue number2
DOIs
StatePublished - Jul 15 2008

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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