Abstract
The phenol-based total protein extraction method is unique in that water-soluble components such as polyphenolic compounds and nucleic acids can be easily removed. Thus, total protein is free from contaminants and allows for high quality two–dimensional gel electrophoresis. The phenol-based extraction of total protein was used in various lily organs and may likely apply to other plants whose content of polyphenolics is high (Note 1). An additional advantage of this extraction method is that nucleic acids can be easily removed and thus, avoid adverse effects of nucleic acids on protein resolution in the gel. This method is modified from that of Hurkman and Tanaka (1986).
Keywords: Protein extratction, Phenol, Plant
Materials and Reagents
Equipment
Procedure
Representative data
Figure 1. Anther-specificity analysis of lily protein by SDS-PAGE. Total protein (40 μg) was isolated from various organs of L. longiflorum, separated by 12% SDS-PAGE and stained with Coomassie blue. 1, root; 2, stem; 3, leaf; 4, tepal; 5, filament; 6, anther; 7, pistil; M, marker. Figure 2. 2D-PAGE analysis of lily pollen protein. Total protein (1.5 mg) was isolated from pollen of L. longiflorum, separated by 2D-PAGE and stained with Coomassie blue. Marker proteins are indicated in the left.
Recipes
Notes
Acknowledgments
The protocol is mainly modified from that of Hurkman and Tanaka (1986). We appreciate their original contribution.
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.