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Chromatographic amino acid analysis of protein hydrolysed in polyacrylamide after removal of ammonia

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Summary

A simple method for the removal of NH3 from amino acids is presented. The method is based on a cation-exchange resin from which amino acids are eluted with NH4OH. The eluent is then removed under reduced pressure. The method allows the ninhydrin-based detection of amino acids after hydrolysis of stained protein bands in polyacrylamide gels. This was previously not possible since NH3 produced by the hydrolysis of polyacrylamide interferes with the ninhydrin-detection of basic amino acids. The method should also be applicable to the detection of amino acids with o-phthalaldehyde.

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Abbreviations

OPA:

o-phthalaldehyde

SDS:

sodium dodecylsulfate

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Authors and Affiliations

  1. Department of Plant Pathology, and Agricultural Research Service, USDA, University of Wisconsin, 53706, Madison, WI, USA

    E. E. Farmer, T. F. Uchytil, J. P. Helgeson & R. D. Durbin

Authors
  1. E. E. Farmer
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  2. T. F. Uchytil
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  3. J. P. Helgeson
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  4. R. D. Durbin
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Farmer, E.E., Uchytil, T.F., Helgeson, J.P. et al. Chromatographic amino acid analysis of protein hydrolysed in polyacrylamide after removal of ammonia. Chromatographia 23, 115–117 (1987). https://doi.org/10.1007/BF02312885

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  • DOI: https://doi.org/10.1007/BF02312885

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