A combined radiolabelling and silver staining technique for improved visualisation, localisation, and identification of proteins separated by two-dimensional gel electrophoresis

PROTEOMICS ◽  
2001 ◽  
Vol 1 (3) ◽  
pp. 370-376 ◽  
Author(s):  
Jules A. Westbrook ◽  
Jun X. Yan ◽  
Robin Wait ◽  
Michael J. Dunn
1987 ◽  
Vol 50 (1) ◽  
pp. 17-21 ◽  
Author(s):  
Huw A. John ◽  
Ian F. Purdom

SummaryThe protein spots representing the haptoglobin α1F, α1Sand α2chains in two-dimensional gels of human plasma samples representative of the six common haptoglobin phenotypes were identified by comparing their position with those of purified haptoglobin and distinguished from other spots in the vicinity by comparison with plasma with undetectably low levels of haptoglobin. Silver staining indicated that the α1Fchain was represented by one spot in subtypes 1F-1F, 1F-1S and 1F-2, the α1Schain by three spots in subtypes 1S-1S, 1F-1S and 1S-2 and the α2chain by six spots in 1F-2, 1S-2 and 2–2


1985 ◽  
Vol 54 (03) ◽  
pp. 626-629 ◽  
Author(s):  
M Meyer ◽  
F H Herrmann

SummaryThe platelet proteins of 9 thrombasthenic patients from 7 families were analysed by high resolution two-dimensional gel electrophoresis (HR-2DE) and crossed immunoelectrophoresis (CIE). In 7 patients both glycoproteins (GPs) IIb and Ilia were absent or reduced to roughly the same extent. In two related patients only a trace of GP Ilb-IIIa complex was detected in CIE, but HR-2DE revealed a glycopeptide in the position of GP Ilia in an amount comparable to type II thrombasthenia. This GP Ilia-like component was neither recognized normally by anti-GP Ilb-IIIa antibodies nor labeled by surface iodination. In unreduced-reduced two-dimensional gel electrophoresis two components were observed in the region of GP Ilia. The assumption of a structural variant of GP Ilia in the two related patients is discussed.


2015 ◽  
Vol 22 (12) ◽  
pp. 1066-1075 ◽  
Author(s):  
Adriana Magalhães ◽  
Rayner Queiroz ◽  
Izabela Bastos ◽  
Jaime Santana ◽  
Marcelo Sousa ◽  
...  

Author(s):  
Fatemeh Nasri ◽  
Maryam Zare ◽  
Mehrnoosh Doroudchi ◽  
Behrouz Gharesi-Fard

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.


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