Background Protein extracts obtained from cells or tissues often contain interfering substances, which must be removed for preparing high-quality protein samples [ 1 ]. Material and methods Plant materials Maize Zea mays L. Download: PPT.
Fig 1. Acetone precipitation One-step acetone precipitation was performed as described recently [ 17 ]. Fig 2. Evaluation of the modified method First, we made a comprehensive comparison of protein yields and resolution in 2-DE by the modified and classical methods. Table 1.
Comparison of protein yield and spot number in 2DE between the two methods. Fig 3. Comparison of 2DE protein profiles of maize embryo proteins extracted using two methods. Table 2. The identification of the differential extracted proteins in maize using the two methods. Fig 4. Comparison of 2DE profiles of maize mesocotyl proteins extracted using two methods.
Supporting information. S1 Fig. S2 Fig. Comparison of 2DE protein profiles of maize root proteins extracted using two methods. S3 Fig. Comparison of 2DE protein profiles of maize leaf proteins extracted using two methods. S4 Fig. S1 Table. Comparison of spot variations in 2DE gels of maize mesocotyl. S2 Table. Comparison of spot variations in 2DE gels of maize embryo. References 1. Trichloroacetic acid TCA precipitation of proteins. Cold Spring Harb Protoc. Optimizing protein extraction from plant tissues for enhanced proteomics analysis.
J Sep Sci. Front Plant Sci. Total protein extraction with TCA-acetone. Methods Mol. Sample extraction techniques for enhanced proteomic analysis of plant tissues. Protein extraction from plant tissues for 2DE and its application in proteomic analysis. Proteomics ; — Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat-seedling proteins.
Electrophoresis ; 7: 52—4. View Article Google Scholar 8. Comparison of protein precipitation methods for various rat brain structures prior to proteomic analysis. A critical evaluation of sample extraction techniques for enhanced proteomic analysis of recalcitrant plant tissues.
Sample preparation for two-dimensional gel electrophoresis. Acetone precipitation of proteins and the modification of peptides. J Proteome Res. A universal and rapid protocol for protein extraction from recalcitrant plant tissues for proteomic analysis. Electrophoresis ; —6. A simple protein extraction method for proteomic analysis of mahogany Swietenia macrophylla embryos.
Plant Omics. View Article Google Scholar Changes in the 2-DE protein profile during zygotic embryogenesis in the Brazilian Pine Araucaria angustifolia. As always, if you have any tips, hints, or suggestions to share, please speak up in the comments section!
Next up: protein concentration via chromatography. Part 1: Semi-permeable membranes. Part 2: Protein precipitation. Part 3: Chromatography. Has this helped you? Then please share with your network. You must be logged in to post a comment. This site uses Akismet to reduce spam. Learn how your comment data is processed. Facebook Twitter LinkedIn More. Written by Jode Plank. Log in to Reply. Dave on December 7, at am. Great article thanks a lot for posting. Dr Amanda Welch on December 7, at am.
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