Quantification of proteins in the subnanogram and nanogram range: comparison of the AuroDye, FerriDye, and India ink staining methods.

Analytical Biochemistry
K W LiJ Joosse

Abstract

The usefulness of three sensitive dyes, AuroDye, FerriDye, and India ink, for the quantification of proteins and peptides bound to nitrocellulose paper has been assessed. In general, the staining intensity varies linearly with the logarithm of protein concentrations. The detection limit of small peptides (Mr less than 5000) is higher than that of large peptides and proteins, but the sensitivity is independent of the molecular weight. Oligopeptides of four or less amino acids either stain with very high detection limits or do not stain at all. The detection limit of proteins stained by AuroDye is approximately 1 ng, and in a number of cases even lower. The useful range for quantification of proteins extends to around 100 ng. The FerriDye and India ink staining methods are less sensitive and can be used to quantify proteins over a wide nanogram range. Among the methods tested, the India ink staining method has the highest protein to protein variation in sensitivity.

References

Dec 1, 1975·Analytical Biochemistry·C E Stauffer
Aug 1, 1983·Analytical Biochemistry·K Hancock, V C Tsang

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Citations

Sep 23, 2000·Electrophoresis·P MajercakovaG Peltre
Feb 1, 1991·Journal of Biochemical and Biophysical Methods·R J RouwenhorstJ P van Dijken
Oct 19, 2013·Analytical Biochemistry·Ghanshyam D HedaRajiv P Heda
Sep 23, 2006·Proteomics·Ingrid MillerElisabetta Gianazza
May 18, 1999·Analytical Biochemistry·T Morçöl, A Subramanian
Apr 28, 1995·Journal of Chromatography. a·P J Wirth, A Romano

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