An improved, luminescent europium-based stain for detection of electroblotted proteins on nitrocellulose or polyvinylidene difluoride membranes

Electrophoresis
C KemperW F Patton

Abstract

SYPRO Rose Plus protein blot stain is an improved europium-based metal chelate stain for the detection of proteins on nitrocellulose and poly(vinylidene difluoride) (PVDF) membranes. Staining is achieved without covalently modifying the proteins. The stain may be excited with a 254 nm (UV-C), 302 nm (UV-B), or 365 nm (UV-A) light source and displays a sharp emission maximum at 612 nm. The emission peak has a full width at half-maximum of only 8 nm. The stain exhibits exceptional photostability, allowing long exposure times for maximum sensitivity. Since the dye is composed of a europium complex, it has a long emission lifetime, potentially allowing time-resolved detection, greatly reducing background fluorescence. Proteins immobilized to a nitrocellulose or PVDF membrane by electroblotting, dot-blotting, or vacuum slot-blotting are incubated with SYPRO Rose Plus protein blot stain for 15-30 min. Membranes are rinsed briefly, visualized with UV epi-illumination and the luminescence of the europium dye is measured using a 490 nm long-pass or 625 +/- 15 nm band-pass filter in combination with a conventional photographic or charge-coupled device (CCD) camera system. Alternatively, the dye may be visualized using a xenon-arc illumin...Continue Reading

Citations

May 11, 2011·Analytical and Bioanalytical Chemistry·A K Hagan, T Zuchner
Apr 2, 2008·Applied Microbiology and Biotechnology·Anna Maria SwistowskaBernd Hofer
Dec 18, 2013·Proteomics·Christian P MoritzEckhard Friauf
Sep 23, 2006·Proteomics·Ingrid MillerElisabetta Gianazza
Oct 21, 2004·Analytical Biochemistry·Guang-Chao ZhaoZhou-Sheng Yang
May 26, 2004·Analytical Biochemistry·Babu S AntharavallyA Krishna Mallia
May 10, 2011·Analytical Biochemistry·Tracey RaeJeffrey Fishpaugh

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