Research Article

Analysis of the efficiency of post-electrphoretic protein staining using colloidal Coomassie Blue G-250

Alicja Dratwa-Chałupnik , Adam Lepczyński, Małgorzata Ożgo, Agnieszka Herosimczyk, Katarzyna Michałek, Magdalena Niemcewicz, Anna Pałyszka, Wiesław Skrzypczak

West Pomeranian University of Technology, Szczecin, Poland

1. Acta Sci. Pol. Zootechnica, 14(2), 2015, 67‒76

Abstract. Constant improvements in the technique for the electrophoretic separation of proteins have resulted in development of more sensitive and less time-consuming staining methods. Despite the fact that fluorescent dyes are highly sensitive, classical methods such as silver or coomassie blue still represent very popular staining techniques. However, information concerning the comparison of the same stain but used in different protocols are sparse. Based on the results of many different authors it is difficult to clearly assess the efficiency of staining protocol. Therefore, present study was aimed at evaluating (in respect of sensitivity, simplicity and time of staining procedure and also the degree of environmental hazard) two different staining procedures with Coomassie Blue G-250 for proteins in polyacrylamide gels. A comparison of two CBB-based procedures for staining proteins showed that modified protocol of Pink is much more efficient in protein detection. This staining technique also takes less time to carry out and it is less toxic as it requires no methanol when compared to the Hoving protocol.

Keywords: 1-DE, 2-DE, CBB G-250, Hoving protocol, Pink protocol