http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
Solubilization of Proteins from Human Lymph Node Tissue and Two-Dimensional Gel Storage
De Marqui, Alessandra Bernadete Trovo,Vidotto, Alessandra,Polachini, Giovana Mussi,De Mattos Bellato, Claudia,Cabral, Hamilton,Leopoldino, Andreia Machado,De Gois Filho, Jose Francisco,Fukuyama, Erica Korean Society for Biochemistry and Molecular Biol 2006 Journal of biochemistry and molecular biology Vol.39 No.2
In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT(dithiothreitol) and 0.2% carrier ampholytes; (b) 5M urea, 2M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate), 40mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7M urea, 2M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.
Solubilization of Proteins from Human Lymph Node Tissue and Two-Dimensional Gel Storage
Alessandra Bernadete Trovo De Marqui,Alessandra Vidotto,Giovana Mussi Polachini,Claudia De Mattos Bellato,Hamilton Cabral,Andreia Machado Leopoldino,Jose Francisco De Gois Filho,Erica Erina Fukuyama,F 한국생활환경학회 2006 BMB Reports Vol.39 No.2
Functional activities of the Tsh protein from avian pathogenic Escherichia coli (APEC) strains
Renata K. T. Kobayashi,Luis Carlos J. Gaziri,Marilda C. Vidotto 대한수의학회 2010 Journal of Veterinary Science Vol.11 No.4
The temperature-sensitive hemagglutinin (Tsh) expressed by strains of avian pathogenic Escherichia (E.) coli (APEC) has both agglutinin and protease activities. Tsh is synthesized as a 140 kDa precursor protein, whose processing results in a 106 kDa passenger domain (Tshs) and a 33 kDa β-domain (Tshβ). In this study, both recombinant Tsh (rTsh) and supernatants from APEC, which contain Tshs (106 kDa), caused proteolysis of chicken tracheal mucin. Both rTsh (140 kDa) and pellets from wild-type APEC, which contain Tshβ (33 kDa), agglutinated chicken erythrocytes. On Western blots, the anti-rTsh antibody recognized the rTsh and 106 kDa proteins in recombinant E. coli BL21/pET 101-Tsh and in the supernatants from APEC grown at either 37oC or 42oC. Anti-rTsh also recognized a 33 kDa protein in the pellets from APEC13 cultures grown in either Luria-Bertani agar, colonization factor antigen agar, or mucin agar at either 26oC, 37oC, or 42oC, and in the extracts of outer membrane proteins of APEC. The 106 kDa protein was more evident when the bacteria were grown at 37oC in mucin agar, and it was not detected when the bacteria were grown at 26oC in any of the culture media used in this study. Chicken anti-Tsh serum inhibited hemagglutinating and mucinolytic activities of strain APEC13 and recombinant E. coli BL21/pET101-Tsh. This work suggests that the mucinolytic activity of Tsh might be important for the colonization of the avian tracheal mucous environment by APEC.
Solubilization of Proteins from Human Lymph Node Tissue andTwo-Dimensional Gel Storage
?rica Erina Fukuyama,Patr?cia Maluf Cury,Mario Sergio Palma,Alessandra Bernadete Trov? de Marqui,Giovana Mussi Polachini,Hamilton Cabral,Eloiza Helena Tajara,Alessandra Vidotto,Cl?udia de Mattos Bella 한국생화학분자생물학회 2006 BMB Reports Vol.39 No.2