Journal of capillary electrophoresis最新文献_第5页

A study of an erythrocyte membrane protein that contributes to inhibition of agglutination of feline erythrocytes in glucose solution 一种在葡萄糖溶液中抑制猫红细胞凝集的红细胞膜蛋白的研究

Journal of capillary electrophoresis Pub Date : 2010-01-01 DOI: 10.2198/JELECTROPH.54.9

K. Namikawa, Yumi Sato, T. Maruo, F. Sunaga, K. Sakaguchi, J. Suzuki

{"title":"A study of an erythrocyte membrane protein that contributes to inhibition of agglutination of feline erythrocytes in glucose solution","authors":"K. Namikawa, Yumi Sato, T. Maruo, F. Sunaga, K. Sakaguchi, J. Suzuki","doi":"10.2198/JELECTROPH.54.9","DOIUrl":"https://doi.org/10.2198/JELECTROPH.54.9","url":null,"abstract":"Due to the negative charges on their surface membrane, erythrocytes usually do not agglutinate each other in vivo. However, when mixed with 5% glucose solution in a test tube, some feline erythrocytes exhibit agglutination. To investigate the reasons for this phenomenon, we extracted erythrocyte membrane proteins from these cells and subjected them to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In samples in which agglutination did not occur, a band was seen at the 36-kDa position, whereas in samples showing agglutination, this band disappeared or has lower intensity. The 36-kDa position corresponds to glycophorin A in the human erythrocyte membrane, but no immunochemical cross-reaction with this band was seen. We therefore conducted mass spectrometry in order to investigate the composition, and found a partial amino acid sequence, His-Ile-Thr-Ser-Tyr-Pro-Glu-Thr-His-Glu-Gly. Furthermore, although no protein showing this sequence was found in any database, this protein was confirmed to be an acidic glycoprotein, therefore it is thought to be a glycophorin-like molecule in the feline erythrocyte membrane that could contribute to inhibition of agglutination.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"1 1","pages":"9-12"},"PeriodicalIF":0.0,"publicationDate":"2010-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88706146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Analysis of plasma protein in newborn rats treated with diethylstilbestrol by using rapid micro two-dimensional electrophoresis 二乙烯雌酚处理新生大鼠血浆蛋白的快速微双向电泳分析

Journal of capillary electrophoresis Pub Date : 2009-12-01 DOI: 10.2198/JELECTROPH.53.51

J. Suzuki, Shogo Ujiie, Ryosuke Kira, K. Sakaguchi, Yoichi Kakuno, M. Honda

{"title":"Analysis of plasma protein in newborn rats treated with diethylstilbestrol by using rapid micro two-dimensional electrophoresis","authors":"J. Suzuki, Shogo Ujiie, Ryosuke Kira, K. Sakaguchi, Yoichi Kakuno, M. Honda","doi":"10.2198/JELECTROPH.53.51","DOIUrl":"https://doi.org/10.2198/JELECTROPH.53.51","url":null,"abstract":"We examined the effect of diethylstilbestrol(DES) administration on newborn male rats by rapid micro two-dimensional polyacrylamide gel electrophoresis (RM2D-PAGE) under non-denaturing/denaturing conditions and a 10-17% second-dimensional gel. We began by visually analyzing the changes at each week over time (weeks 1-9) in plasma protein spots from the same individual DES-administered male rats. We found the tendency for mercaptoalbumin (reduced form) spots to increase in week 1 in DES-administered rats. We also analyzed the quantitative alterations in α2u-globulin (AUG), a biomarker of endocrine disruptors, and found a two-week delay in expression in DES-administered rats in comparison to those not given DES (control rats). We then investigated the reproducibility of these results by using samples taken from multiple male rats in weeks 5, 7 and 9 to perform electrophoretic analysis three times for each individual, and observed the changes in AUG. AUG production was not seen in week 5 in the control rats, but was present at weeks 7 and 9. In DES-administered rats, expression was not observed at week 5 or 7, but was present at week 9. In addition, we identified the spots of AUG by Western blotting after electrophoresis. We consequently confirmed that AUG production delayed by two weeks in DES-administered rats, in comparison with control rats, which suggests that the effect of either a decline in AUG synthesis function or its depletion results in stunting of the reproductive organs, which was confirmed through autopsy.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"53 1","pages":"51-56"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73996304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Toward high performance western blotting using vacuum-driven system and laser scanner; comparison between two signal detection methods based on chemiluminescent and immunofluorescent imaging 利用真空驱动系统和激光扫描仪进行高效免疫印迹的研究化学发光与免疫荧光成像两种信号检测方法的比较

Journal of capillary electrophoresis Pub Date : 2009-12-01 DOI: 10.2198/JELECTROPH.53.63

Chisato Harada, H. Tajima, S. Hirohashi, T. Kondo

引用次数: 0

Miniaturized two-dimensional gel electrophoresis of high-molecular-weight proteins using low-concentration multifilament-supporting gel for isoelectric focusing 使用低浓度多丝支持凝胶进行等电聚焦的高分子量蛋白质的小型化二维凝胶电泳

Journal of capillary electrophoresis Pub Date : 2009-12-01 DOI: 10.2198/JELECTROPH.53.57

T. Odake, T. Saheki, K. Kamezawa, Kozue Miyata, H. Takiguchi, H. Hotta, Onda Koki, Yasuaki Kojima, Jinxiang Li, K. Tsunoda

{"title":"Miniaturized two-dimensional gel electrophoresis of high-molecular-weight proteins using low-concentration multifilament-supporting gel for isoelectric focusing","authors":"T. Odake, T. Saheki, K. Kamezawa, Kozue Miyata, H. Takiguchi, H. Hotta, Onda Koki, Yasuaki Kojima, Jinxiang Li, K. Tsunoda","doi":"10.2198/JELECTROPH.53.57","DOIUrl":"https://doi.org/10.2198/JELECTROPH.53.57","url":null,"abstract":"A miniaturized two-dimensional gel electrophoresis (2-DE) system was constructed with a multifilament-supporting (MFS) gel as a first-dimensional gel for isoelectric focusing (IEF). The MFS gel was 1 mm in diameter, had a multifilament yarn of acrylic as a gel-support, and could provide low-concentration (2.5%T) polyacrylamide with sufficient mechanical strength, resulting in fast IEF, and clear 2-D spots due to the relatively large pore and improved transfer efficiency from the first-dimensional gel to the second-dimensional one of especially high-molecular-weight (HMW) proteins. In this study, by using the MFS gel of 4 cm in length as a miniaturized IEF gel, three colored proteins were separated in 8 min, which was about twice faster than using the MFS gel of standard length (7 cm). The second-dimensional gel was also miniaturized to be 55 mm wide, 40 mm long and 0.75 mm thick. The low-concentration MFS gel after IEF was easily transferred onto the top of the miniaturized second-dimensional gel with a tweezers. By using the miniaturized 2-DE system, the HMW size marker was separated in 30 min by IEF and in 25 min by native-polyacrylamide gel electrophoresis (Native-PAGE), which was at least twice faster than by using the standard size 2-DE system using MFS gel. This miniaturized 2-DE system could be expected as a useful separation method for fast protein diagnosis and screening.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"77 1","pages":"57-61"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88560341","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Characterization of acidic and neutral streptolysin O 酸性和中性溶血素O的表征

Journal of capillary electrophoresis Pub Date : 2009-09-01 DOI: 10.2198/JELECTROPH.53.45

J. Suzuki

{"title":"Characterization of acidic and neutral streptolysin O","authors":"J. Suzuki","doi":"10.2198/JELECTROPH.53.45","DOIUrl":"https://doi.org/10.2198/JELECTROPH.53.45","url":null,"abstract":"We previously confirmed that crude streptolysin O (SLO) consists of heterogeneous components with different isoelectric points (pIs) and hemolytic efficiency, as determined by the slope of the titration curve. The two preparations of SLO having different pI distributions were further characterized in this research. The pIs of the major components of the two SLO preparations are around pH 6 (acidic) and 7.5 (neutral). Although the neutral SLO was shown to be more hydrophobic than the acidic SLO based on the behavior on hydrophobic interaction chromatography, the capacities of the two types to bind to erythrocytes did not differ. On the other hand, the neutral SLO destroyed liposomes to some extent, whereas the acidic SLO did not, even at concentrations of 100 HD50 (50% hemolytic dose), suggesting that the mode of membrane damage by the two types of SLO are different. Furthermore, electron microscopic observation indicated that the acidic SLO preferably formed an irregular-shaped complex and semicircular pores on erythrocyte membranes, in contrast to the complete pore formation by the neutral SLO. Thus, we clarified that the two hemolytic forms of SLO have different pore formation efficiencies based on liposome membrane damage and electron micrographs of ghosts.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"7 1","pages":"45-50"},"PeriodicalIF":0.0,"publicationDate":"2009-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85634786","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

The parameters related to small dense low-density lipoprotein evaluated by non-denaturing polyacrylamide gel electrophoresis method in dyslipidemic subjects : with reference to the Japanese guideline for prevention of atherosclerotic cardiovascular diseases 非变性聚丙烯酰胺凝胶电泳法评价血脂异常受试者小密度低密度脂蛋白相关参数:参照日本预防动脉粥样硬化性心血管疾病指南

Journal of capillary electrophoresis Pub Date : 2009-09-01 DOI: 10.2198/JELECTROPH.53.39

K. Tsuzaki, K. Kotani, Kazunori Yamada, Y. Sano, Yukiyo Matsuoka, Kentaro Okazaki, Toshiyuki Yamada, A. Shimatsu, N. Sakane

{"title":"The parameters related to small dense low-density lipoprotein evaluated by non-denaturing polyacrylamide gel electrophoresis method in dyslipidemic subjects : with reference to the Japanese guideline for prevention of atherosclerotic cardiovascular diseases","authors":"K. Tsuzaki, K. Kotani, Kazunori Yamada, Y. Sano, Yukiyo Matsuoka, Kentaro Okazaki, Toshiyuki Yamada, A. Shimatsu, N. Sakane","doi":"10.2198/JELECTROPH.53.39","DOIUrl":"https://doi.org/10.2198/JELECTROPH.53.39","url":null,"abstract":"To prevent the development of atherosclerotic diseases, the Japanese guideline for prevention of atherosclerotic cardiovascular diseases (released from Japan Atherosclerosis Society in 2007) is clinically referred. In addition, the importance of evaluating not only the level but also the quality of low-density lipoprotein (LDL) has been advocated. Small dense LDL (sdLDL) is regarded as representation of the quality of LDL. With the LipoprintTM LDL subfraction system, utilizing non-denaturing polyacrylamide gel electrophoresis method which can evaluate several sdLDL-related parameters such as the area percentage of sdLDL, mean LDL particle size (mean LDL-PS), relative mobility in the form of the relative flotation rate (Rf), and LDL size-based phenotypic patterns. We attempted to assess the Japanese guideline for prevention of atherosclerotic cardiovascular diseases in terms of sdLDL-related parameters. We studied with 46 asymptomatic and cardiovascular disease-free dyslipidemic subjects (23 men, aged 30-58 years olds; 23 women, 31-58 years olds), who were within the primary prevention level. From the increased number of categories in the guideline, the following trends were observed; the smaller LDL-PS (p=0.002), the larger of area percentage of sdLDL (p=0.042), an increase in Rf (p=0.068), and an increase in the presence of pattern B (atherogenic phenotype) (p=0.031). These results suggested that, with the LipoprintTM system, the usage of Japanese guideline may clinically reflect well the sdLDL-related parameters, in particular the mean LDL-PS, the area percentage of sdLDL and the pattern B.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"124 1","pages":"39-43"},"PeriodicalIF":0.0,"publicationDate":"2009-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79395552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Proteome analysis of silkworm 2. Hemolymph 家蚕蛋白质组学分析2。血淋巴

Journal of capillary electrophoresis Pub Date : 2009-06-01 DOI: 10.2198/JELECTROPH.53.27

H. Kajiwara, A. Imamaki, Masatoshi Nakamura, K. Mita, Q. Xia, M. Ishizaka

引用次数: 10

Proteome analysis of silkworm 3. Malpighian tube 家蚕蛋白质组学分析马氏管

Journal of capillary electrophoresis Pub Date : 2009-06-01 DOI: 10.2198/JELECTROPH.53.33

H. Kajiwara, A. Imamaki, Masatoshi Nakamura, K. Mita, Q. Xia, M. Ishizaka

引用次数: 2

Proteome analysis of silkworm 1. Fat body 家蚕蛋白质组学分析脂肪体

Journal of capillary electrophoresis Pub Date : 2009-06-01 DOI: 10.2198/JELECTROPH.53.19

H. Kajiwara, A. Imamaki, Masatoshi Nakamura, K. Mita, Q. Xia, M. Ishizaka

{"title":"Proteome analysis of silkworm 1. Fat body","authors":"H. Kajiwara, A. Imamaki, Masatoshi Nakamura, K. Mita, Q. Xia, M. Ishizaka","doi":"10.2198/JELECTROPH.53.19","DOIUrl":"https://doi.org/10.2198/JELECTROPH.53.19","url":null,"abstract":"Proteome analysis of silkworm (Bombyx mori L.) fat bodies was performed on the fifth-instar day-3 larva stage and the expression of proteins separated on two-dimensional polyacrylamide gels was tracked from the fourth-instar day-1 larva stage to the stage before adult moth. Proteins on the 2D-gels were excised manually and treated with 4-vinylpyridine for alkylation. Each spot was analyzed by capillary high-performance liquid chromatography coupled with ion-trap mass spectrometry after proteolysis using trypsin. In the previous report1), the amino acid sequence database obtained from the Drosophila genome was used for the protein identification. This analysis used the amino acid sequence data elucidated from the silkworm genome. Additionally, protein expression in fat bodies was compared with that at other stages and in other tissues described in the silkworm proteome database (http://kaiko2ddb.dna.affrc.go.jp/cgi-bin/search_2DDB.cgi). Several unidentified proteins from the previous report1) were identified with high Mascot scores. Induction and reduction of proteins during metamorphosis were observed as changes in spot concentration on sequential 2D-gels on the web.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"9 1","pages":"19-26"},"PeriodicalIF":0.0,"publicationDate":"2009-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88631712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7

The usefulness of electrophoretic analysis of urinary retinol-binding protein and retinol-binding protein 4 in type 2 diabetes mellitus patients 2型糖尿病患者尿视黄醇结合蛋白和视黄醇结合蛋白4电泳分析的意义

Journal of capillary electrophoresis Pub Date : 2009-03-01 DOI: 10.2198/JELECTROPH.53.7

K. Kanamori, M. Kameko, H. Kitamura, Junichi Nishimaki, K. Shiba, Kenji Sato

引用次数: 0