Appendix

APPENDIX 3J Detection and Quantitation of Radiolabeled Proteins and DNA in Gels and Blots

  1. Daniel Voytas,
  2. Ning Ke

Published Online: 1 NOV 2002

DOI: 10.1002/0471142735.ima03js50

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Voytas, D. and Ke, N. 2002. Detection and Quantitation of Radiolabeled Proteins and DNA in Gels and Blots. Current Protocols in Immunology. 50:3J:A.3J.1–A.3J.10.

Author Information

  1. Iowa State University, Ames, Iowa

Publication History

  1. Published Online: 1 NOV 2002
  2. Published Print: AUG 2002

Abstract

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.