Direct amino acid analysis of proteins electroblotted onto polyvinylidene difluoride membrane from sodium dodecyl sulfate-polyacrylamide gel.

Analytical Biochemistry
S Nakagawa, T Fukuda

Abstract

The band of appropriate proteins (basic pancreatic trypsin inhibitor, soybean trypsin inhibitor, interleukin 2, and human leukocyte interferon alpha A) on a polyvinylidene difluoride (PVDF) membrane, which was electroblotted from sodium dodecyl sulfate (SDS)-polyacrylamide gel and then stained with Coomassie blue R-250, was cut out and directly hydrolyzed in HCl in the presence of thioglycolic acid for amino acid analysis. The analytical values agreed with those expected with recoveries of 29-47%, except that the value for tryptophan was very low or scarcely detected. This method was applied to the identification of human growth hormone (hGH) in a partially purified preparation. The amino acid composition of the band corresponding to about 2 micrograms of hGH agreed with the theoretical values. These results indicate that the band on the PVDF membrane can be directly hydrolyzed for amino acid analysis and that the method can be used for partially purified proteins separated using SDS-polyacrylamide gel electrophoresis.

References

Feb 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·J R Benson, P E Hare
Jul 31, 1985·Biochemical and Biophysical Research Communications·K KatoA Kakinuma
Apr 30, 1971·Science·A Chrambach, D Rodbard
Apr 29, 1969·Biochemical and Biophysical Research Communications·H Matsubara, R M Sasaki
Jan 1, 1964·Analytical Biochemistry·D RACUSEN, N CALVANICO
Dec 1, 1964·Analytical Biochemistry·A L KOEN, C R SHAW

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Citations

Aug 1, 1997·Electrophoresis·I Humphery-SmithW P Blackstock
May 1, 1990·Biochimica Et Biophysica Acta·R E BrownK J Hyland
Jun 7, 1996·Journal of Chromatography. a·J X YanD F Hochstrasser
Oct 3, 1993·Biochimica Et Biophysica Acta·K KamemuraT Takahashi
Feb 9, 1996·Biochimica Et Biophysica Acta·K KamemuraT Takahashi
Jan 23, 1999·Journal of Chromatography. a·M Fountoulakis, H W Lahm
Dec 4, 2012·Journal of Bioscience and Bioengineering·Kyoshiro NonakaSeiji Ogo
Nov 26, 2009·Current Protocols in Protein Science·Shane M Rutherfurd, G Sarwar Gilani
Jan 1, 1998·Bioscience, Biotechnology, and Biochemistry·S Kabuto, K Muramoto

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