Journal•ISSN: 0031-9082
Seibutsu Butsuri Kagaku
Japanese Electrophoresis Society
About: Seibutsu Butsuri Kagaku is an academic journal. The journal publishes majorly in the area(s): Gel electrophoresis & Polyacrylamide gel electrophoresis. Over the lifetime, 847 publications have been published receiving 704 citations.
Topics: Gel electrophoresis, Polyacrylamide gel electrophoresis, Gel electrophoresis of proteins, Isoelectric focusing, Two-dimensional gel electrophoresis
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TL;DR: The master 2-D gel pattern with spot identification numbers and the database of age-dependent protein alteration were presented and a new project has started in the institute for establishing my own age-related protein database of human diploid fibroblasts upon a2-D PAGE technique.
Abstract: Organismic aging appears in the results of functional alteration of tissue-constituent cells. The replicative life span of normal human diploid fibroblasts in cell culture has been realized as a typical model of the research on mitotic cell aging. From the above premises, a new project has started in our institute for establishing my own age-related protein database of human diploid fibroblasts upon a 2-D PAGE technique. The protocol of 2-D PAGE with horizontal IPG-IEF and vertical SDS-PAGE was standardized including whole procedures from cell harvest through spot quantification, since the reproducibility of 2-D gel pattern depended on the procedure. The master 2-D gel pattern of human diploid fibroblasts was generated by merging skin fibroblast-specific spots into the standard 2-D gel pattern of lung fibroblasts in an image analyzing software, because fibroblasts showed tissue specificity. The master 2-D gel pattern with spot identification numbers and the database of age-dependent protein alteration were presented on my home page for free access through the Internet computer network.
36 citations
26 citations
TL;DR: The determination of nitrite and nitrate in human blood plasma by capillary zone electrophoresis (CZE) was carried out using 750 mM sodium chloride containing 5% NICE-Pak OFM Anion-BT as running buffer, at a potential of 20 kV, with on-column UV detection at 214 nm.
Abstract: キャピラリー電気泳動を用いて血漿 nitrite, nitrate 濃度を測定した. キャピラリーは65cm×75μmの fused-silica 型のものを用い, 電圧は20kV, 検出波長は214nmとした. 泳動バッファーは5%NICE-Pak OFM Anion-BTを含む750mM NaClとした. 血液は, 採血後1000g, 5分間遠心分離して, 血漿を-80℃で保存した. 血漿は測定前にフィルターし, 濾液の20μlを使用した.nitrite 濃度と泳動ピークの面積は正の相関を示し, その回帰式はy=1425+8261xでr2=0.999であった. nitrate 濃度と泳動ピークの面積も同様に正の相関を示し, その回帰式はy=1417+8416xでr2=0.999であった. 41名の健康成人の静脈血 nitrite, nitrate 濃度はそれぞれ, 0.15±0.07mg/l, 3.20±1.60mg/lであった. 敗血症, 亜硝酸ナトリウム投与, クモ膜下出血後の3症例の経過中に nitrite, nitrate を測定し, 病態により血漿中 nitrate が変化することがわかった. 以上より, キャピラリー電気泳動による血漿中 nitrite, nitrate 測定は, 集中治療患者のモニターとして有用と思われる.
23 citations
17 citations
TL;DR: Addition of 1M thiourea in combination with 5M urea to the medium for agarose gel made it possible to be used with ease on micro-scale as well as on macro-scale electrophoresis.
Abstract: A procedure was established for micro-two-dimensional gel electrophoresis with agarose gels in the first dimension. Agarose (Agarose IEF, Pharmacia) was previously found to be suitable for the analysis of high molecular weight proteins with a wide range of isoelectric points, but rather difficult to be handled for micro-scale electrophoresis. However, addition of 1M thiourea in combination with 5M urea to the medium for agarose gel made it possible to be used with ease on micro-scale as well as on macro-scale. Since thiourea is a potent protein solubilizing reagent, almost all amounts of proteins including structural proteins were presented on the respective positions on two-dimensional gels. This method is applicable to other experiments with high molecular weight proteins to be analyzed in a wide range of isoelectric points.
16 citations