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Optica Publishing Group
  • Applied Spectroscopy
  • Vol. 43,
  • Issue 2,
  • pp. 349-350
  • (1989)

Indirect Fluorometric Detection in Gel Electrophoresis

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Abstract

Polyacrylamide gel electrophoresis separates biomolecules by virtue of their inherent charge or size, or a combination of the two. This widely used bioanalytical technique has seen applications in areas such as the separation of complex mixtures of proteins, gene sequencing and mapping, and clinical diagnosis. The method displays unmatched resolution of macromolecules, yet still utilizes detection methods which are slow and labor-intensive. Commonly used methods such as Coomassie Blue and silver staining can become complicated and may require twenty-four hours or longer to allow one to visualize the analyte. Autoradiography offers high sensitivity (10-100 pg) for the detection of radio-labelled analytes, but requires exposure times of hours or days. A sensitive, yet rapid, quantitation method for electrophoresis would be quite useful. This paper demonstrates the feasibility of employing indirect fluorometry as a rapid and potentially highly sensitive detection scheme for gel electrophoresis.

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