Abstract
Unlike the traditional methods for staining of polyacrylamide gels (e.g., Coomassie blue, silver stain) whose purpose is to dye the protein bands (1,2), the purpose of reverse staining (RS) methods is to leave protein bands unstained and dye the rest of the gel instead. RS methods are usually based on the precipitation of some metal salt all along the gel, except in the positions where proteins are. The contrast between protein bands and background, and the sensitivity of a particular RS method, will depend on both the intensity of the selective precipitation and the color of the precipitate.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Garfin, D. E. (1990) One-dimensional gel electrophoresis, in Guide to Protein Purification (Deutscher, M. P., ed.), Academic, San Diego, CA, pp. 425–441.
Merril, C. R. (1990) Gel-staining techniques, in Guide to Protein Purification (Deutscher, M. P., ed.), Academic, San Diego, CA, pp. 477–488.
Lee, C., Levin, A., and Branton, D. (1987) Copper staining: a five-minute protein stain for sodium dodecyl sulfate-polyacrylamide gels. Anal. Biochem. 166, 308–312.
Dzandu, J. K., Johnson, J. F., and Wise, G. E. (1988) Sodium dodecyl sulfate-gel electrophoresis staining of polypeptides using heavy metal salts. Anal. Biochem. 174, 157–167.
Fernández-Patrón, C., Castellanos-Serra, L., and Rodríguez, P. (1992) Reverse staining of sodium dodecyl sulfate polyacrylamide gels by imidazole-zinc salts: sensitive detection of unmodified proteins. Biotechniques 12, 564–573.
Ortiz, M. L., Calero, M., Fernández-Patrón, C., Castellanos, L., and Méndez, E. (1992) Imidazole-SDS-Zn reverse staining of proteins in gels containing or not SDS and microsequence of individual unmodified electroblotted proteins. FEBS Lett. 296, 300–304.
Ferreras, M., Gavilanes, J. G., and García-Segura, J. M. (1993) A permanent Zn2+ reverse staining method for the detection and quantification of proteins in polyacrylamide gels. Anal. Biochem. 213, 206–212.
Pliss, G. B. and Zabezhinskii, M. A. (1970) Carcinogenic properties of orthotoluidine (3,3′-dimethylbenzidine). J. Natl. Cancer Inst. 45, 283–289.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Stoll, V. S. and Blanchard, J. S. (1990) Buffers: principles and practice, in Guide to Protein Purification (Deutscher, M. P., ed.), Academic, San Diego, CA, pp. 24–38.
Fernández-Patrón, C., Hardy, E., Sosa, A., Seoane, J., and Castellanos, L. (1995) Double staining of Coomassie blue-stained polyacrylamide gels by imidazole sodium dodecyl sulfate-zinc reverse staining: sensitive detection of Coomassie blue-undetected proteins. Anal. Biochem. 224, 263–269.
Fernández-Patrón, C., Calero, M., Rodríguez Collazo, P., Garcia, J. R., Madrazo, J., Musacchio, A., Soriano, F., Estrada, R., Frank, R., Castellanos-Serra, L. R., and Méndez, E. (1995) Protein reverse staining: high-efficiency microanalysis of unmodified proteins detected on electrophoresis gels. Anal. Biochem. 224, 203–211.
Burriel, F., Arribas, S., Lucena, F., and Hernández, J. (1985) Química Analítica Cualitativa, Paraninfo, Madrid, p. 695.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1996 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
García-Segura, J.M., Ferreras, M. (1996). Zn2+-Reverse Staining of Proteins in Polyacrylamide Gels. In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-60327-259-9_29
Download citation
DOI: https://doi.org/10.1007/978-1-60327-259-9_29
Publisher Name: Humana Press
Print ISBN: 978-0-89603-338-2
Online ISBN: 978-1-60327-259-9
eBook Packages: Springer Book Archive