Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
12
pubmed:dateCreated
2006-12-22
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.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0897-7151
pubmed:author
pubmed:issnType
Print
pubmed:volume
23
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1794-801
pubmed:dateRevised
2007-12-3
pubmed:meshHeading
pubmed:year
2006
pubmed:articleTitle
beta-tubulin is a more suitable internal control than beta-actin in western blot analysis of spinal cord tissues after traumatic injury.
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.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Research Support, N.I.H., Extramural