EN
Coomassie Brilliant Blue R-250 (CBBR-250) is
the most frequently used dye stain in protein research.
However, relatively poor sensitivity and long application
durations are the major limitations of this stain. In the
present study, we have investigated the effect of pre-stain
methanol washing on the sensitivity of the conventional
CBB staining method. Concentrations of methanol ranging
from 5 to 30% were prepared in 5% acetic acid. Pre-stain
washing of SDS-gels using acetic acid (5%) and methanol
were assessed at various timings and temperatures. Our
results demonstrate that pre-stain washing by methanol
greatly improves the sensitivity of the CBB stain and
facilitates the visualization of low molecular weight
components present in a complex muscle protein,
actomyosin. The outcome of this study also revealed
duration of pre-stain washing and temperature as other
factors that determine the sensitivity of CBB staining
method. Therefore, it is suggested that pre-stain washing
with methanol may improve the sensitivity of CBB stain in
SDS-PA gels.