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.

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

Proteome analysis of Leishmania (Viannia) braziliensis by two-dimensional gel electrophoresis and mass spectrometry

2007 ◽  
Vol 154 (1) ◽  
pp. 6-21 ◽  
Author(s):  
Patricia Cuervo ◽  
Jose Batista de Jesus ◽  
Magno Junqueira ◽  
Leila Mendonça-Lima ◽  
Luis Javier González ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gel Electrophoresis ◽  
Proteome Analysis ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis

scholarly journals Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis

2011 ◽  
Vol 5 ◽  
pp. BCBCR.S6263 ◽  
Author(s):  
Olena Zakharchenko ◽  
Christina Greenwood ◽  
Louise Alldridge ◽  
Serhiy Souchelnytskyi
Keyword(s):  
Gel Electrophoresis ◽  
Breast Tissue ◽  
Protein Extraction ◽  
Small Sample Size ◽  
Small Sample ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Main Challenge ◽  
Optimized Protocol ◽  
Separation Of Proteins

Proteomics is a highly informative approach to analyze cancer-associated transformation in tissues. The main challenge to use a tissue for proteomics studies is the small sample size and difficulties to extract and preserve proteins. The choice of a buffer compatible with proteomics applications is also a challenge. Here we describe a protocol optimized for the most efficient extraction of proteins from the human breast tissue in a buffer compatible with two-dimensional gel electrophoresis (2D-GE). This protocol is based on mechanically assisted disintegration of tissues directly in the 2D-GE buffer. Our method is simple, robust and easy to apply in clinical practice. We demonstrate high quality of separation of proteins prepared according to the reported here protocol.

scholarly journals Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction

2010 ◽  
Vol 2010 ◽  
pp. 1-9 ◽  
Author(s):  
Magnus Centlow ◽  
Stefan R. Hansson ◽  
Charlotte Welinder
Keyword(s):  
Signal Transduction ◽  
Gel Electrophoresis ◽  
Protein Separation ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Proteome Analysis ◽  
Apolipoprotein A1 ◽  
Two Dimensional ◽  
2D Page ◽  
Nutritional Effect

The human placenta is a difficult tissue to work with using proteomic technology since it contains large amounts of lipids and glycogen. Both lipids and glycogen are known to interfere with the first step in the two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), the isoelectric focusing. In order to gain the best possible protein separation on 2D-PAGE, an optimized sample preparation protocol for placental proteins was developed. Two different buffers, urea/CHAPS and Hepes, were used for solubilization in combination with six different precipitation methods. The removal of glycogen from the samples by centrifugation was crucial for the final proteome maps. Solubilization with urea/CHAPS in combination with dichloromethane/methanol or acidified acetone proved to be the best precipitation procedures. When applied to clinical placenta samples apolipoprotein A1 was found to be accumulated in the preeclamptic placenta, where it may either have a nutritional effect or act as a modifier of signal transduction.

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