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

Display

Search in Pubmed

Author: Poon, HF
Author: Crawford, F
Related works

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 6 Enlarged

Fig. 6:

Match rates of two 2D gel-based redox proteomics methodologies. Match rates of 2D Western blot (2DWB), 2D FHA labeling (2DFL), 2DWB + manual assisted matching (MA) and 2DFL + MA using PDQuest. Data represent the average matching rates of 2DWB, 2DFL, 2DWB + MA and 2DFL + MA. Measured values are normalized to the 2DWB values. Error bar indicate the SEM of the match rate, * p < 0.005. 2DWB, match rates between 2D Western blot of protein carbonyl and Coomassie gel; 2DFL, match rates between 2D gel of fluorescently labeled carbonyl and Coomassie gel; 2DWB + MA, match rates between 2D Western blot of protein carbonyl and Coomassie gel after manual assistance using spots from Fig. 3B as landmark references; 2DFL + MA, Match rates between 2D gel of fluorescently labeled carbonyl and Coomassie gel after manually assistance using the spot in Fig. 4A as landmark references.

Disclaimer/Legal Copyright Privacy Policy