An electroelution apparatus for sequential transfer of sodium dodecyl sulfate-proteins into agarose and mass spectrometric identification of Li-Na-dodecyl sulfate-proteins from solubilized agarose

• Published in: Electrophoresis 2004-04, Vol.25 (7-8), p.966-969
• Main Authors: Buzás, Zsuzsanna, Antal, József, Gilligan, John J., Backlund, Peter S., Yergey, Alfred L., Chrambach, Andreas
• Format: Article
• Language: English
• Subjects: Agarose solubilization; Electrophoresis, Polyacrylamide Gel - instrumentation; Gel band transfer apparatus; Lithium - chemistry; Matrix-assisted laser desorption/ionization-mass spectrometry; Proteins - isolation & purification; Sepharose - chemistry; Sequential electroelution; Sodium - chemistry; Solubility; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.200305797

Separated protein bands are sequentially electrophoresed into low melting agarose plugs distributed in an apparatus of original design along the surface of a plastic drum. The rotation of the drum is synchronized to migration of electrophoretic bands to receive each band individually. Agarose plugs are dissolved enzymatically for transfer into the mass spectrometer. One μL of the agarose solution containing 1 pmol of each of three lithium and natrium salts of dodecyl sulfate (Li‐Na‐DS)‐proteins were applied to matrix‐assisted laser desorption/ionization‐mass spectrometry (MALDI‐MS) without any prepurification. It yields a signal indistinguishable from that obtained in the absence of agarose.