An appropriate loading control for western blot analysis in animal models of myocardial ischemic infarction
| Autor: | Sheng Hu, Xin Nie, Y. James Kang, Wenjing Zhang, Fulai Xue, Chen Li |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
SDS sodium dodecyl sulfate PVDF polyvinylidene fluoride Myocardial ischemia Biophysics Western blot Housekeeping proteins Anterior Descending Coronary Artery Biochemistry Total protein Andrology lcsh:Biochemistry EDTA ethylene diamine tetraacetic acid 03 medical and health sciences 0302 clinical medicine Ischemic infarction medicine lcsh:QD415-436 Myocardial infarction lcsh:QH301-705.5 PAGE polyacrylamide gel electrophoresis Glyceraldehyde 3-phosphate dehydrogenase biology medicine.diagnostic_test business.industry CBB Coomassie brilliant blue medicine.disease Molecular biology Staining LAD left anterior descending coronary artery TBS Tris-buffered saline 030104 developmental biology lcsh:Biology (General) GAPDH glyceraldehyde-3-phosphate dehydrogenase HKP housekeeping protein H&E hematoxylin and eosin biology.protein TBST TBS with Tween Loading control Ligation business 030217 neurology & neurosurgery Research Article |
| Zdroj: | Biochemistry and Biophysics Reports, Vol 12, Iss C, Pp 108-113 (2017) Biochemistry and Biophysics Reports |
| ISSN: | 2405-5808 |
| Popis: | 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. Highlights • Loading control for WB has been studied in animal models of myocardial ischimia. • GAPDH is a reliable loading control in Rhesus monkeys model of myocardial ischemia. • Total protein level is the best loading control for western blot in animal models. |
| Databáze: | OpenAIRE |
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