Measuring the absolute abundance of the Smad transcription factors using quantitative immunoblotting.
In the age of systems biology, biologists seek to quantify the absolute number of molecules in experimentally treated samples. Immunoblotting remains a technique of choice for assessing the relative differences between the protein levels in different samples. Here we discuss how to exploit immunoblotting for estimating the number of Smad transcription factor molecules per cell. We focus on describing the calculations needed to analyze the data. Our methods are generally applicable to the quantification of other cellular proteins.
Clarke
David C
DC
Department of Chemistry and Biochemistry, University of Colorado at Boulder, Boulder, CO, USA.
Liu
Xuedong
X
D002457:Cell Extracts; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D012394:Rosaniline Dyes / Q000378:metabolism; D051785:Smad Proteins / Q000378:metabolism*; D012984:Software; D013194:Staining and Labeling; D013223:Statistics as Topic; D016212:Transforming Growth Factor beta / Q000494:pharmacology
D002457:Cell Extracts; D012394:Rosaniline Dyes; D051785:Smad Proteins; D016212:Transforming Growth Factor beta; C004692:coomassie Brilliant Blue