Background: Protein extraction from plant tissues is a great challenge since they contain
low amount of proteins and rich in proteases, secondary metabolites and oxidative enzymes. Besides,
dealing with limited amount of starting material would pose another challenge in getting high
protein yield. The aim of this study is to determine the best protein extraction protocol for recalcitrant
tissues of banana (Musa sp.) specific for non-reducing one dimensional SDS-PAGE analysis.
Methods: Three protein extraction protocols were compared which include TCA-acetone, phenol
and phosphate buffer saline (PBS) using small amount of starting material. Modifications were done
to each protocol to suit non-reducing SDS-PAGE analysis and subsequent downstream processes.
Results: Of the three protocols compared, TCA-acetone protocol gave the highest yield and quality
of protein extract from only a small amount of starting material based on protein quantification and
non-reducing one dimensional SDS-PAGE analysis compared to phenol and phosphate buffer saline
Conclusion: Even though various protein extraction protocols have been developed to date, there is
no one-size-fits-all when it comes to different types of plant tissues from varieties of plant species.
TCA-acetone protocol was found to be most effective in extracting protein from banana shoot and
root tissues even for small amount of starting material in absence of reducing agent.