Consulta: subjectFacets:"2-DE"
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Data/hora: 01/06/2024 06:43:58

Method optimization for proteomic analysis of soybean leaf: improvements in identification of new and low-abundance proteins
Provedor de dados: 74
Autores: Mesquita,Rosilene Oliveira; Soares,Eduardo de Almeida; Barros,Everaldo Gonçalves de; Loureiro,Marcelo Ehlers.
The most critical step in any proteomic study is protein extraction and sample preparation. Better solubilization increases the separation and resolution of gels, allowing identification of a higher number of proteins and more accurate quantitation of differences in gene expression. Despite the existence of published results for the optimization of proteomic analyses of soybean seeds, no comparable data are available for proteomic studies of soybean leaf tissue. In this work we have tested the effects of modification of a TCA-acetone method on the resolution of 2-DE gels of leaves and roots of soybean. Better focusing was obtained when both mercaptoethanol and dithiothreitol were used in the extraction buffer simultaneously. Increasing the number of washes...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Proteomics; Glycine max; 2-DE; Protein extraction; Leaf proteome.
Ano: 2012
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200017

A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots
Provedor de dados: 74
Autores: Rodrigues,Elisete Pains; Torres,Adalgisa Ribeiro; Batista,Jesiane Stefânia da Silva; Huergo,Luciano; Hungria,Mariangela.
Sample preparation is a critical step in two-dimensional gel electrophoresis (2-DE) of plant tissues. Here we describe a phenol/SDS procedure that, although greatly simplified, produced well-resolved and reproducible 2-DE profiles of protein extracts from soybean [Glycine max (L.) Merril] roots. Extractions were made in three replicates using both the original and simplified procedure. To evaluate the quality of the extracted proteins, ten spots were randomly selected and identified by mass spectrometry (MS). The 2-DE gels were equally well resolved, with no streaks or smears, and no significant differences were observed in protein yield, reproducibility, resolution or number of spots. Mass spectra of the ten selected spots were compared with database...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: 2-DE; Protein extraction; Proteomics; Soybean roots; Symbiosis.
Ano: 2012
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200016

Proteomic data show an increase in autoantibodies and alpha-fetoprotein and a decrease in apolipoprotein A-II with time in sera from senescence-accelerated mice
Provedor de dados: 56
Autores: Guo,S.J.; Qi,C.H.; Zhou,W.X.; Zhang,Y.X.; Zhang,X.M.; Wang,J.; Wang,H.X.
We evaluated changes in levels by comparing serum proteins in senescence-accelerated mouse-prone 8 (SAMP8) mice at 2, 6, 12, and 15 months of age (SAMP8-2 m, -6 m, -12 m, -15 m) to age-matched SAM-resistant 1 (SAMR1) mice. Mice were sacrificed, and blood was analyzed by 2-dimensional electrophoresis combined with mass spectrometry. Five protein spots were present in all SAMP8 serum samples, but only appeared in SAMR1 samples at 15 months of age except for spot 3, which also showed a slight expression in SAMR1-12 m sera. Two proteins decreased in the sera from SAMP8-2 m, -6 m, and -12 m mice, and divided into 2 spots each in SAMP8-15 m sera. Thus, the total number of altered spots in SAMP8 sera was 7; of these, 4 were identified as Ig kappa chain V region...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: SAMP8; Serum proteins; 2-DE; Mass spectrometry; M-T413.
Ano: 2013
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2013000500417

Cardiac protein expression patterns are associated with distinct inborn exercise capacity in non-selectively bred rats
Provedor de dados: 56
Autores: Ribeiro,L.P.; Freitas-Lima,L.C.; Naumann,G.B.; Meyrelles,S.S.; Lunz,W.; Pires,S.F.; Andrade,H.M.; Carnielli,J.B.T.; Figueiredo,S.G.
In the present study, we successfully demonstrated for the first time the existence of cardiac proteomic differences between non-selectively bred rats with distinct intrinsic exercise capacities. A proteomic approach based on two-dimensional gel electrophoresis coupled to mass spectrometry was used to study the left ventricle (LV) tissue proteome of rats with distinct intrinsic exercise capacity. Low running performance (LRP) and high running performance (HRP) rats were categorized by a treadmill exercise test, according to distance run to exhaustion. The running capacity of HRPs was 3.5-fold greater than LRPs. Protein profiling revealed 29 differences between HRP and LRP rats (15 proteins were identified). We detected alterations in components involved in...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Inborn aerobic capacity; Proteomics; 2-DE; MS; Heart proteins.
Ano: 2018
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2018000300609

Assessment of four different detergents used to extract membrane proteins from Xylella fastidiosa by two-dimensional electrophoresis
Provedor de dados: 58
Autores: Di Ciero,Luciana; Bellato,Cláudia de M.; Meinhardt,Lyndel W.; Ferrari,Fernanda; Castellari,Rafael R.; Marangoni,Sérgio; Novello,José C.
Four different detergents, ASB 14, SB 3-10, CHAPS and Triton X100, were utilized to determine the optimal detergent for the solubilization of membrane proteins from the phytopathogenic bacterium Xylella fastidiosa. These proteins were differentially solubilized in distinct buffers containing the detergent and subjected to bidimensional electrophoresis within the non-linear pH range of 3-10. The detergents ASB 14 and SB 3-10 were the most effective revealing 221 and 157 spots, respectively. CHAPS and Triton X100 were less effective and revealed only 72 and 43 spots, respectively. MALDI-TOF tryptic peptide mass fingerprinting of 18 excised proteins from the ASB 14 treatment revealed that 83% were membrane proteins and that the theoretical efficiency of...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: X. fastidiosa; 2-DE; Extraction; Proteome; Solubilization.
Ano: 2004
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822004000200018

Challenges in proteome analyses of tropical plants
Provedor de dados: 61
Autores: Balbuena,Tiago S.; Dias,Leonardo L. C.; Martins,Mariana L. B.; Chiquieri,Tatiana B.; Santa-Catarina,Claudete; Floh,Eny I. S.; Silveira,Vanildo.
Genome sequencing of various organisms allow global analysis of gene expression, providing numerous clues on the biological function and involvement in the biological processes studied. Proteomics is a branch of molecular biology and biotechnology that has undergone considerable development in the post-genomic era. Despite the recent significant advancements in proteomics techniques, still there is much to be improved. Due to peculiarities to the plant kingdom, proteomics approaches require adaptations, so as to improve efficiency and accuracy of results in plants. Data generated by proteomics can substantially contribute to the understanding and monitoring of plant physiological events and development of biotechnological strategies. Especially for...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: 2-DE; Mass spectrometry; Recalcitrant species; Proteomics; Unknown genome.
Ano: 2011
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1677-04202011000200001

Two-dimensional gel electrophoretic protein profile analysis during seed development of Ocotea catharinensis: a recalcitrant seed species
Provedor de dados: 61
Autores: Dias,Leonardo L.C.; Balbuena,Tiago S.; Silveira,Vanildo; Santa-Catarina,Claudete; Schevchenko,Andrej; Floh,Eny I. S.
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...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: 2-DE; Comparative proteomics; Embryo development; Storage protein; Lauraceae.
Ano: 2010
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1677-04202010000100003