Improved conditions for fluorescent staining of proteins with 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid in SDS-PAGE.
A simple and sensitive fluorescent staining method for the detection of proteins in SDS-PAGE, namely IB (improved 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid) stain, is described. Non-covalent hydrophobic probe 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid was applied as a fluorescent dye, which can bind to hydrophobic sites in proteins non-specifically. As low as 1 ng of protein band can be detected briefly by 30 min washing followed by 15 min staining without the aiding of stop or destaining step. The sensitivity of the new presented protocol is similar to that of SYPRO Ruby, which has been widely accepted in proteomic research. Comparative analysis of the MS compatibility of IB stain and SYPRO Ruby stain allowed us to address that IB stain is compatible with the downstream of protein identification by PMF.
Cong
Wei-Tao
WT
College of Pharmacy & Research Institute of Drug Development, Chonnam National University, Gwangju, South Korea.
Hwang
Sun-Young
SY
Jin
Li-Tai
LT
Choi
Jung-Kap
JK
D000817:Anilino Naphthalenesulfonates / Q000737:chemistry*; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D016014:Linear Models; D009942:Organometallic Compounds / Q000737:chemistry*; D011506:Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D009942:Organometallic Compounds; D011506:Proteins; C515601:Sypro Ruby; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate)