Detection and quantitation of radiolabeled proteins and DNA in gels and blots.
This appendix presents procedures for visualizing and quantitating radiolabeled proteins or DNA separated by polyacrylamide gel electrophoresis or affixed to filter membranes. Autoradiography is the most common method by which this is accomplished; X-ray film is the traditional recording medium. Film images can be analyzed by densitometry as described to obtain a quantitative measure of the radioactivity in the sample. The use of X-ray films for autoradiography, however, suffers from lack of sensitivity and limited linear range. Protocols are provided for overcoming these limitations using fluorography and intensifying screens, both of which enhance the radioactive signal. Sensitivity and linear ranges of measurement can be greatly extended by using a phosphor imaging system, which also makes it much faster and easier to quantify radioactive samples.
Voytas
Daniel
D
Iowa State University, Ames, Iowa.
Ke
Ning
N
Iowa State University, Ames, Iowa.
D001345:Autoradiography; D004247:DNA / Q000032:analysis*; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D015151:Immunoblotting; D007553:Isotope Labeling; D011506:Proteins / Q000032:analysis*; D011856:Radiographic Image Enhancement; D011868:Radioisotopes; D014962:X-Ray Film
D011506:Proteins; D011868:Radioisotopes; D004247:DNA