Gel-based detection of protein S-nitrosothiols has relied on the biotin-switch method. This method attempts to replace the nitroso group with a biotin label to allow detection and isolation of S-nitrosated proteins and has been used extensively in the literature. This chapter describes a modification of this method that differs from the original in two major ways. First, it uses a combination of copper ions and ascorbate to achieve selective reduction of the S-nitrosothiol. Second, it replaces the biotin label with fluorescent cyanine dyes in order to directly observe the modified proteins in-gel and perform comparative studies using difference gel electrophoresis analysis in two dimensions. © 2008 Elsevier Inc. All rights reserved.
CITATION STYLE
Kettenhofen, N. J., Wang, X., Gladwin, M. T., & Hogg, N. (2008). In-Gel Detection of S-Nitrosated Proteins Using Fluorescence Methods. In Methods in Enzymology (Vol. 441, pp. 53–71). Academic Press Inc. https://doi.org/10.1016/S0076-6879(08)01204-4
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