Immunoblotting conditions for human hemoglobin chains

Yusuke Suzuki, Yoshihiko Takeda, Tohru Ikuta

Research output: Contribution to journalArticle

16 Citations (Scopus)

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

Fingerprint

Globins
Immunoblotting
Hemoglobins
Collodion
Glutaral
Membranes
Molecular Weight
Molecular weight
Proteins
Metallothionein
Calmodulin
Gene expression
Gene Expression

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Immunoblotting conditions for human hemoglobin chains. / Suzuki, Yusuke; Takeda, Yoshihiko; Ikuta, Tohru.

In: Analytical Biochemistry, Vol. 378, No. 2, 15.07.2008, p. 218-220.

Research output: Contribution to journalArticle

Suzuki, Yusuke ; Takeda, Yoshihiko ; Ikuta, Tohru. / Immunoblotting conditions for human hemoglobin chains. In: Analytical Biochemistry. 2008 ; Vol. 378, No. 2. pp. 218-220.
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