scholarly journals A robust protein extraction method for two dimensional electrophoresis of silkworm proteins

2013 ◽  
Vol 04 (04) ◽  
pp. 584-589
Author(s):  
Lvgao Qin ◽  
Haifeng Shi ◽  
Hengchuan Xia ◽  
Yong Wang ◽  
Liang Chen ◽  
...  
Keyword(s):  
Extraction Method ◽  
Protein Extraction ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis

Protein extraction for two-dimensional electrophoresis from olive leaf, a plant tissue containing high levels of interfering compounds

2003 ◽  
Vol 24 (14) ◽  
pp. 2369-2375 ◽  
Author(s):  
Wei Wang ◽  
Monica Scali ◽  
Rita Vignani ◽  
Antonia Spadafora ◽  
Elisabetta Sensi ◽  
...  
Keyword(s):  
Plant Tissue ◽  
Protein Extraction ◽  
Two Dimensional ◽  
Olive Leaf ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis

Protein extraction fromPhoenix dactylifera L. leaves, a recalcitrant material, for two-dimensional electrophoresis

2008 ◽  
Vol 29 (2) ◽  
pp. 448-456 ◽  
Author(s):  
Sonia Gómez-Vidal ◽  
Manuel Tena ◽  
Luis Vicente Lopez-Llorca ◽  
Jesús Salinas
Keyword(s):  
Protein Extraction ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis ◽  
Recalcitrant Material

scholarly journals Optimization of an Efficient Protein Extraction Protocol Compatible with Two-Dimensional Electrophoresis and Mass Spectrometry from Recalcitrant Phenolic Rich Roots of Chickpea (Cicer arietinum L.)

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Moniya Chatterjee ◽  
Sumanti Gupta ◽  
Anirban Bhar ◽  
Sampa Das
Keyword(s):  
Mass Spectrometry ◽  
Protein Extraction ◽  
Sodium Dodecyl ◽  
Good Resolution ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Extraction Protocol ◽  
Phenol Extraction ◽  
Spot Number ◽  
Dimensional Electrophoresis

Two-dimensional electrophoresis and mass spectrometry are undoubtedly two essential tools popularly used in proteomic analyses. Utilization of these techniques however largely depends on efficient and optimized sample preparation, regarded as one of the most crucial steps for recovering maximum amount of reliable information. The present study highlights the optimization of an effective and efficient protocol, capable of extraction of root proteins from recalcitrant phenolic rich tissues of chickpea. The widely applicable TCA-acetone and phenol-based methods have been comparatively evaluated, amongst which the latter appeared to be better suited for the sample. The phenol extraction-based method further complemented with sodium dodecyl sulphate (SDS) and pulsatory treatments proved to be the most suitable method represented by greatest spot number, good resolution, and spot intensities. All the randomly selected spots showed successful identification when subjected to further downstream MALDI-TOF and MS/MS analyses. Hence, the information obtained collectively proposes the present protein extraction protocol to be an effective one that could be applicable for recalcitrant leguminous root samples.

scholarly journals Evaluation of six different protocols for protein extraction from rice young panicles by two-dimensional electrophoresis

2020 ◽  
Author(s):  
Xiaoping Huang ◽  
Hui-wen Zhou
Keyword(s):  
Protein Concentration ◽  
Protein Extraction ◽  
Image Resolution ◽  
Plant Tissues ◽  
Two Dimensional ◽  
Protein Patterns ◽  
Molecular Weight Range ◽  
Two Dimensional Electrophoresis ◽  
Phenol Extraction ◽  
Dimensional Electrophoresis

Abstract Background: For most reported proteomics approaches, protein extraction are of crucial importance for optimal results. However, extraction of protein from plant tissues still exist great challenges due to low protein content and many secondary metabolites that prominently interfering with isoelectric focusing. Up to now, no attempts are focused on comparison of protein extraction from rice young panicles.Methods: To establish a higher efficiency protein extraction protocol suited for two-dimensional electrophoresis in rice young panicles, six protocols for protein preparation were evaluated in terms of protein concentration, the molecular weight range of protein, gel image resolution, the number of protein spots: 1) Phenol extraction; 2) Mg/Nonidet P-40 (NP-40) extraction; 3) Tris-Base/acetone extraction; 4) SDS extraction; 5) trichloroacetic acid (TCA)/acetone/phenol extraction; 6) TCA/acetone precipitation.Results: The result explicitly demonstrated that TCA/acetone/phenol method provided a high-enhanced protein extraction efficacy from rice young panicles than other protocols in terms of the protein concentration (9.79±0.23 SD), the most comprehensive proteins (10 KDa to 150 KDa), the maximum number of protein spots (450±53 SD), the greater gel image resolution and spot abundance. In addition, these methods also generated remarkably differential 2-DE protein patterns. Twenty-nine of 30 visible differentially extracted proteins were identified by MS analysis and were divided into eight categories. Prediction for protein subcellular localization and grand average of hydropathy (GRAVY) analysis showed that certain special proteins respectively necessitate different extraction methods due to different physicochemical properties of each protocol.Conclusions: Overall, this paper will facilitate to provide a cornerstone of comparative proteomic analysis from rice young panicles, including other complicated plant tissues.

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