Improving image analysis in 2DGE-based redox proteomics by labeling protein carbonyl with fluorescent hydroxylamine

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Author: Poon, HF
Author: Crawford, F
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Methods Article

Improving image analysis in 2DGE-based redox proteomics by labeling protein carbonyl with fluorescent hydroxylamine

H. Fai Poon¹, Laila Abdullah¹, Jon Reed¹, Sarah M Doore¹, Cyndi Laird¹, Venkat Mathura¹, Michael Mullan¹ and Fiona Crawford¹

¹ Roskamp Institute. Sarasota, FL 34243. USA.

* To whom correspondence should be addressed: H. Fai Poon, Roskamp Institute. 2040 Whitfield Ave., Sarasota, FL 34243. USA. Phone: +941 752 2949. Fax: +941 752 2948. Email: faipoon@rfdn.org

Biol. Proced. Online 2007;9:65-72. doi:10.1251/bpo134
Submitted: August 31, 2007; Accepted: December 09, 2007; Published: December 19, 2007.

Indexing terms: electrophoresis, gel, two-dimensional; oxidation-reduction; protein processing, post-translational.

Abbreviations: FHA, fluorescent hydroxylamine; IEF, isoelectric focusing; MALDI, Matrix-assisted laser desorption/ionization.

Figure 4 Enlarged

Fig. 4:

Detection of oxidized bovine serum albumin (BSA) and egg albumin using FHA. (A) SDS-PAGE of oxidized, reduced, and control BSA samples incubated with FHA. (B) Gel showing BSA treated with various concentrations of H₂O₂. (C) Linear increase in fluorescence as a function of concentration of H₂O₂ added.

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