An appropriate loading control for western blot analysis in animal models of myocardial ischemic infarction

Xin Nie, Chen Li, Sheng Hu, Fulai Xue, Yujian Kang, Wenjing Zhang

Research output: Contribution to journalArticle

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Abstract

An appropriate loading control is critical for Western blot analysis. Housekeeping proteins (HKPs), such as β-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and β-tubulin, are commonly used to normalize protein expression. But HKP expression can be impacted by certain experimental conditions, such as ischemic myocardial infarction. This study was undertaken to look for an appropriate loading control for western blot analysis of ischemic myocardium. Myocardial ischemic infarction was induced by left anterior descending coronary artery (LAD) ligation in Rhesus monkeys and C57BL/6 mice. The heart tissue samples from different areas and time points after surgery were subjected to western blot or gel staining. The level of β-actin, GAPDH, β-tubulin, and total protein were tested. The total protein level was consistent in all groups, whereas the protein level of β-tubulin and β-actin were different in all groups. However, the protein level of GAPDH was stable in the Rhesus monkey model. We concluded that total protein was the most appropriate internal control in different stages of myocardial ischemic disease of various animal models. GAPDH is a reliable internal control only for ischemic myocardium of Rhesus monkey.

Original languageEnglish (US)
Pages (from-to)108-113
Number of pages6
JournalBiochemistry and Biophysics Reports
Volume12
DOIs
StatePublished - Dec 1 2017
Externally publishedYes

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Animals
Animal Models
Western Blotting
Myocardial Infarction
Glyceraldehyde-3-Phosphate Dehydrogenases
Proteins
Tubulin
Macaca mulatta
Actins
Housekeeping
Myocardium
Cardiomyopathies
Inbred C57BL Mouse
Surgery
Ligation
Coronary Vessels
Gels
Tissue
Staining and Labeling

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

An appropriate loading control for western blot analysis in animal models of myocardial ischemic infarction. / Nie, Xin; Li, Chen; Hu, Sheng; Xue, Fulai; Kang, Yujian; Zhang, Wenjing.

In: Biochemistry and Biophysics Reports, Vol. 12, 01.12.2017, p. 108-113.

Research output: Contribution to journalArticle

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AU - Zhang, Wenjing

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