Source:http://linkedlifedata.com/resource/pubmed/id/17184189
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
12
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pubmed:dateCreated |
2006-12-22
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pubmed:abstractText |
Western blot is a widely used method for determining specific protein levels. To control and correct for loading error, an internal control is often used. To date, two housekeeping geneâcoded proteins (i.e., beta-actin and beta-tubulin) are widely used as internal controls in the Western blot analysis. However, no information is available concerning the stability of their expressions in response to a traumatic injury to the central nervous system (CNS). If so, their use as an internal control may have a negative impact on data acquisition, analysis, and interpretation. Using Western blot analysis, we demonstrated that spinal cord injury (SCI) induced a significant increase in beta-actin expression which peaked at 7 days post-SCI (2.48-fold). Coefficient of variation (CV) analysis showed that the CV of beta-actin expression was 43.79 +/- 4.67%, significantly higher than that of six loadings from a single sample (6.5 +/- 0.9%, p < 0.01), indicating that increased expression of beta-actin was a result of SCI, instead of a loading error. In contrast, no statistically significant difference was found in beta- tubulin expression following SCI, compared with sham-operated controls. The CV of beta-tubulin expression following SCI was 14.3 beta 3.96%, significantly less than that of the beta-actin expression (43.79 +/- 4.67%; p < 0.01). Taken together, our study suggests that beta-actin whose expression increases following SCI is not a suitable internal control for Western blot analysis of spinal cord tissues following a traumatic injury. In contrast, beta-tubulin, whose expression was not significantly affected by SCI, is a better choice for the internal control.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
0897-7151
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
23
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1794-801
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pubmed:dateRevised |
2007-12-3
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pubmed:meshHeading |
pubmed-meshheading:17184189-Actins,
pubmed-meshheading:17184189-Animals,
pubmed-meshheading:17184189-Blotting, Western,
pubmed-meshheading:17184189-Female,
pubmed-meshheading:17184189-Rats,
pubmed-meshheading:17184189-Rats, Sprague-Dawley,
pubmed-meshheading:17184189-Reference Standards,
pubmed-meshheading:17184189-Reproducibility of Results,
pubmed-meshheading:17184189-Spinal Cord Injuries,
pubmed-meshheading:17184189-Thoracic Vertebrae,
pubmed-meshheading:17184189-Time Factors,
pubmed-meshheading:17184189-Tubulin
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pubmed:year |
2006
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pubmed:articleTitle |
beta-tubulin is a more suitable internal control than beta-actin in western blot analysis of spinal cord tissues after traumatic injury.
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pubmed:affiliation |
Kentucky Spinal Cord Injury Research Center, Departments of Neurological Surgery and Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, Kentucky 40292, USA.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't,
Research Support, N.I.H., Extramural
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