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Coomassie Brilliant Blue R-250 (CBBR-250) isthe most frequently used dye stain in protein research.However, relatively poor sensitivity and long applicationdurations are the major limitations of this stain. In thepresent study, we have investigated the effect of pre-stainmethanol washing on the sensitivity of the conventionalCBB staining method. Concentrations of methanol rangingfrom 5 to 30% were prepared in 5% acetic acid. Pre-stainwashing of SDS-gels using acetic acid (5%) and methanolwere assessed at various timings and temperatures. Ourresults demonstrate that pre-stain washing by methanolgreatly improves the sensitivity of the CBB stain andfacilitates the visualization of low molecular weightcomponents present in a complex muscle protein,actomyosin. The outcome of this study also revealedduration of pre-stain washing and temperature as otherfactors that determine the sensitivity of CBB stainingmethod. Therefore, it is suggested that pre-stain washingwith methanol may improve the sensitivity of CBB stain inSDS-PA gels

Effect of pre-stain methanol washing on the sensitivity of CBBR-250 staining in SDS-polyacrylamide gels