Detection of Proteins on Western Blots Using Chemifluorescence
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Immunodetection of specific proteins that have been immobilized on membrane supports by Western blotting is a widely used protein analysis technique. Traditionally, radioactively labeled antibodies or ligands (usually 125I) were used to probe for specific antigens (1-3). Autoradiographic detection on X-ray film, for example, reveals the location of the antigen with reproducibly high sensitivity and the results are easily quantifiable. However, the need for lengthy exposure times and concerns related to the handling and disposal of radioactive materials have led to the development of nonradio-active techniques for Western blot analysis, for example, using colorimetric and chemi-luminescent detection.
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