Description
The FluoroStain Protein Fluorescent Staining Dye (Red, 1000×) is designed to substitute the common Coomassie Blue protein staining method, offering greater sensitivity and ease of operation. Unlike Coomassie Blue stain, the FluoroStain Protein Fluorescent Staining Dye binds to protein with high specificity, making destaining process an option rather than a requirement. With further reduction of background signals via destaining process, the FluoroStain is capable of achieving detection level parallel to silver staining without specialized imaging equipment (Fig. 1), making it one of the most sensitive dyes available. In addition to its remarkable sensitivity, the FluoroStain Protein Fluorescent Staining Dye (Red, 1000×) brings a more reliable and safer user experience, since the stained gel can be visualized with blue-light illumination, avoiding the risk of skin/ eye damage caused by UV light. For best result, we suggest using B-BOX™ Blue Light LED epi-illuminator to visualize and analyze the gel stained with FluoroStain Protein Fluorescent Staining Dye (Red, 1000×).
The FluoroStain Protein Fluorescent Staining Dye is compatible to the analysis of mass spectra (Fig. 2), i.e. LC-MS/MS, MALDI-TOF, etc.
Fig.1. Comparison of FluoroStain Protein Fluorescent Staining Dye (Red, 1000×) with Silver stain of a 2× serially diluted BSA sample.
Fig.3. Excitation and emission spectrum of FluoroStain Protein Fluorescent Staining Dye (Red, 1000×).
Spectral Characteristics When it is bound with bovine serum albumin (BSA), the fluorescent emission of FluoroStain Protein Fluorescent Staining Dye can be excited by UV and blue light sources, with excitation peaks around 369 and 517 nm and emission at 605 nm (Fig. 3). In absence of BSA, FluoroStain Protein Fluorescent Staining Dye shows ignorable fluorescence as compared with protein-bound form, therefore giving a clear background for photographic analysis.
These spectral characteristics made this fluorescent dye compatible with a wide variety of gel reading facilities, including UV/ blue light epi- and transilluminator, argon laser and mercury-arc lamp excitation gel scanners.
Working Reagent Preparation
1:1000 dilution in 40% ethanol and 2% H3PO4
Storage -20°C ≥ 12 months
Fig.2. Compariosn of MALDI-TOF mass spectra of bovine serum albumin (BSA) stained with FluoroStain Protein Fluorescent Staining Dye (A) and with Coomassie Blue (B). BSA proteins are seperated on SDS-PAGE, stained with fluorescent dye or conventional Coomassie Blue, followed by trypsin digestion in gel, and then analyzed by MALDI-TOF.
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