Evaluation of three types of protein extraction methods for tetraploid black locust (Robinia pseudoacacia L.) phloem tissue proteome analysis by two-dimensional electrophoresis

2015 ◽  
Vol 7 (3) ◽  
pp. 1008-1017 ◽  
Author(s):  
Sheng Zhang ◽  
Ling-Ling Zhang ◽  
Kai-Kai Zhou ◽  
Yu-Jing Liu ◽  
Zhong Zhao
Keyword(s):  
Protein Extraction ◽  
Robinia Pseudoacacia ◽  
Proteome Analysis ◽  
Extraction Methods ◽  
Two Dimensional ◽  
Phloem Tissue ◽  
Two Dimensional Gel Electrophoresis ◽  
Robinia Pseudoacacia L ◽  
Tetraploid Black Locust ◽  
Tissue Proteome

Sample preparation is a decisive step in a two-dimensional gel electrophoresis (2-DE) proteome approach and is essential for good experimental results.

Protein extraction from mature rice leaves for two-dimensional gel electrophoresis and its application in proteome analysis

PROTEOMICS ◽  
2004 ◽  
Vol 4 (7) ◽  
pp. 1903-1908 ◽  
Author(s):  
Nazrul Islam ◽  
M. Lonsdale ◽  
N. M. Upadhyaya ◽  
T. J. Higgins ◽  
H. Hirano ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Protein Extraction ◽  
Proteome Analysis ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Rice Leaves

scholarly journals A comparative method for protein extraction and proteome analysis by two-dimensional gel electrophoresis from banana fruit

2016 ◽  
Vol 2 (1) ◽  
Author(s):  
Giridara Kumar Surabhi
Keyword(s):  
Gel Electrophoresis ◽  
Protein Extraction ◽  
Comparative Method ◽  
Proteome Analysis ◽  
Protein Accumulation ◽  
Banana Fruit ◽  
Two Dimensional ◽  
Fruit Peel ◽  
Two Dimensional Gel Electrophoresis ◽  
Large Matrix

<p>Bananas and plantains are a major staple food and export product in many countries.<strong> </strong>Banana fruit tissues contain large amounts of secondary compounds, polysaccharides and other plant polymers which will interfere with protein extraction for gel-based proteomic analysis. Due to presence of very small amount of proteins (approximately 1%) in a large matrix of fruit tissues, it classified as ‘recalcitrant’.  In this connection, assessment of different protein extraction protocols and extraction of high quality proteins from banana fruit tissues is crucial for successful gel-based proteome analysis.   In this study, two different protein extraction protocols were validated to isolate proteins from banana (cv.Grand Naine) fruit peel and pulp tissues, and proteins were resolved by using SDS-PAGE.  A comparative study showed that phenol based method is effective in extracting proteins over TCA-acetone method. Isolated proteins were further subjected to two-dimensional gel electrophoresis separation and stained with colloidal coomassie blue to visualize protein spots. On average 380 protein spots could be detected on coomassie stained two-dimensional gel and our study clearly demonstrated the differential protein accumulation during pre-climacteric and climacteric stages of banana fruit. <strong></strong></p>

Proteome Analysis of CD4+ T Cells Reveals Differentially Expressed Proteins in Infertile Polycystic Ovary Syndrome Patients

2020 ◽  
Vol 20 ◽  
Author(s):  
Fatemeh Nasri ◽  
Maryam Zare ◽  
Mehrnoosh Doroudchi ◽  
Behrouz Gharesi-Fard
Keyword(s):  
Mass Spectrometry ◽  
T Cells ◽  
T Lymphocytes ◽  
Gel Electrophoresis ◽  
Cd4 T Cells ◽  
Polycystic Ovary ◽  
Proteome Analysis ◽  
Two Dimensional ◽  
Ovary Syndrome ◽  
Two Dimensional Gel Electrophoresis

Background: Polycystic ovary syndrome (PCOS) is the most frequent endocrine disorder affecting 6–7% of premenopausal women. Recent studies revealed that the immune system especially CD4+ T helper cells are important in the context PCOS. Proteome analysis of CD4+ T lymphocytes can provide valuable information regarding the biology of these cells in the context of PCOS. Objective: To investigate immune dysregulation in CD4+ T lymphocytes at the protein level in the context of PCOS using two-dimensional gel electrophoresis (2DE) and mass spectrometry (MS). Methods: In the present study, we applied two-dimensional gel electrophoresis / mass spectrometry to identify proteins differentially expressed by peripheral blood CD4+ T cells in ten PCOS women compared with ten healthy women. Western blot technique was used to confirm the identified proteins. Results: Despite the overall proteome similarities, there were significant differences in the expression of seven spots between two groups (P <0.05). Three proteins, namely phosphatidylethanolamine-binding protein 1, proteasome activator complex subunit 1 and triosephosphate isomerase 1 were successfully identified by Mass technique and confirmed by western blot. All characterized proteins were over-expressed in CD4+ T cells from patients compared to CD4+ T cells from controls (P <0.05). In-silico analysis suggested that the over-expressed proteins interact with other proteins involved in cellular metabolism especially glycolysis and ferroptosis pathway. Conclusion: These findings suggest that metabolic adjustments in CD4+ T lymphocytes, which is in favor of increased glycolysis and Th2 differentiation are important in the context of PCOS.

scholarly journals Two-dimensional gel electrophoretic protein profile analysis during seed development of Ocotea catharinensis: a recalcitrant seed species

2010 ◽  
Vol 22 (1) ◽  
pp. 23-33 ◽  
Author(s):  
Leonardo L.C. Dias ◽  
Tiago S. Balbuena ◽  
Vanildo Silveira ◽  
Claudete Santa-Catarina ◽  
Andrej Schevchenko ◽  
...  
Keyword(s):  
Seed Development ◽  
Protein Identification ◽  
Protein Extraction ◽  
Profile Analysis ◽  
Protein Profile ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Recalcitrant Seed ◽  
And Storage ◽  
Cotyledonary Stage

The aim of the present work was to characterize changes in the protein profile throughout seed development in O. catharinensis, a recalcitrant species, by two-dimensional gel electrophoresis. Protein extraction was undertaken by using a thiourea/urea buffer, followed by a precipitation step with 10% TCA. Comparative analysis during seed development showed that a large number of proteins were exclusively detected in each developmental stage. The cotyledonary stage, which represents the transition phase between embryogenesis and the beginning of metabolism related to maturation, presents the highest number of stage-specific spots. Protein identification, through MS/MS analysis, resulted in the identification of proteins mainly related to oxidative metabolism and storage synthesis. These findings contribute to a better understanding of protein metabolism during seed development in recalcitrant seeds, besides providing information on established markers that could be useful in defining and improving somatic embryogenesis protocols, besides monitoring the development of somatic embryos in this species.

Development of Efficient Protein Extraction Methods for Shotgun Proteome Analysis of Formalin-Fixed Tissues

2007 ◽  
Vol 6 (3) ◽  
pp. 1038-1047 ◽  
Author(s):  
Xiaogang Jiang ◽  
Xinning Jiang ◽  
Shun Feng ◽  
Ruijun Tian ◽  
Mingliang Ye ◽  
...  
Keyword(s):  
Protein Extraction ◽  
Proteome Analysis ◽  
Extraction Methods ◽  
Formalin Fixed

Proteome analysis of glandular parotid and submandibular-sublingual saliva in comparison to whole human saliva by two-dimensional gel electrophoresis

PROTEOMICS ◽  
2006 ◽  
Vol 6 (5) ◽  
pp. 1631-1639 ◽  
Author(s):  
Anke Walz ◽  
Kai Stühler ◽  
Andreas Wattenberg ◽  
Eva Hawranke ◽  
Helmut E. Meyer ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Proteome Analysis ◽  
Human Saliva ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis
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