Departmental Bulletin Paper SYPRO® Ruby を用いたタンパク質染色法の湯浴加熱による迅速・高感度化
Increased Speed and Performance of Protein Staining with SYPRO® Ruby Protein Gel Stain using Hot Water Incubation

尼子, 克己  ,  Amako, Katsumi

8pp.35 - 40 , 2017-03-31 , 仁愛大学
The author presents a novel rapid, low-cost staining protocol (hot water protocol) for proteinsin a polyacrylamide gel using the fluorescent dye SYPRO® Ruby. Compared to the supplierrecommendedprotocol, the hot water protocol showed equivalent dynamic range and linearity, anda lower detection limit. Peptides prepared with in-gel digestion extracted from polyacrylamide gelsstained using the hot water protocol showed a sequence coverage ratio equivalent to that obtainedusing the supplier-recommended protocol, and de novo sequencing could also be carried out withoutissue. In addition, use of the hot water protocol dramatically decreased speckling noises owing toprolonged staining time or storage of the dye solution. These results demonstrate the superiority ofthe hot water protocol over the supplier-recommended protocol.

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