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J Biomol Tech. 2010 September; 21(3 Suppl): S53.
PMCID: PMC2918041

Evaluation of Oriole Fluorescent Gel Stain and Comparison to SYPRO Ruby

V. Thulasiraman and J. Waker, II
Life Science Group, Bio-Rad Laboratories, Hercules, CA, United States

Abstract

RP-102

Fluorescent stains are widely used in gel-based proteomics due to the high sensitivity and wide dynamic range possible with fluorescent detection reagents. Fluorescent metal complexes are particularly well-suited to this task because of their and high stability, wide Stokes shifts, and resistance to photobleaching and quenching, A novel fluorescent metal complex has been developed that can be used to stain SDS-PAGE gels. This metal complex has been formulated into a gel stain called Oriole fluorescent gel stain, which can be applied in a simple one-step staining protocol. This study was undertaken to characterize the staining properties of Oriole stain and evaluate its performance in the proteomics workflow in comparison to SYPRO Ruby, a prominent staining product based on another fluorescent metal complex. Protein mixtures were run on SDS-PAGE. The gels were stained with either Oriole stain or SYPRO Ruby stain and imaged using UV-transillumination. The resulting gel images were evaluated for appearance, sensitivity and protein-to-protein variability. In-gel digestion and MS analysis were applied to excised gel plugs. Two-dimensional gels of complex samples were also stained and evaluated. Optimal staining results with Oriole stain were obtained using a simple protocol that required only immersion of the gel in the stain solution for 90 minutes without fixing or destaining. Staining sensitivity, protein-to-protein consistency and MS compatibility were all equivalent to or better than SYPRO Ruby. Oriole stain fulfills all of the criteria for utility in gel-based proteomics and is uniquely rapid and simple to use.


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