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Detecting Fluorescent Dyes on 1-D Polyacrylamide Gels using CCD-based Technology as Compared to Laser Scanning

Journal article published in 2005 by Florence Goubet, Paul Dupree, Xiaolan Yu, Paru Oatey
This paper was not found in any repository; the policy of its publisher is unknown or unclear.
This paper was not found in any repository; the policy of its publisher is unknown or unclear.

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Preprint: policy unknown
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Abstract

Laser scanners are commonly used to detect some of the newer fluorescent protein stains such as Sypro Ruby (Molecular Probes, Inc., Leiden, The Netherlands), because it provides detection in the nanogram range. However, it does have major drawbacks including being an expensive technology, with limited uses in terms of detecting a range of visible protein stains. Charge-coupled device (CCD) technology offers the advantage of being more flexible in its applications but is often reported as being less sensitive than laser scanning for measuring proteins labeled with fluorescent dyes. In this article, 1-D polyacrylamide protein gels were stained with either Sypro Ruby or Deep Purple (Amersham Biosciences, Little Chalfont, UK), and the gels were visualized by a laser scanner (Typhoon 9400 (Amersham)) set at manufacturer's defined conditions. This was compared with a CCD system (ChemiGenius (Syngene, Cambridge, UK)) using different lighting and filter combinations. The data showed that using the correct lighting and filter combinations, CCD technology can produce results that are as sensitive as laser-based imaging and therefore, offers an inexpensive and accurate alternative.