Journal of capillary electrophoresis最新文献_第4页

Detection of Acidovorax avenae subsp. citrulli using PCR and MALDI-TOF MS 酸霉亚种的检测。用PCR和MALDI-TOF MS鉴定瓜氨酸

Journal of capillary electrophoresis Pub Date : 2012-01-01 DOI: 10.2198/JELECTROPH.56.13

H. Kajiwara, Masatoshi Sato, A. Suzuki

引用次数: 9

Comparison of urinary exosomal protein solubilization methods for two-dimensional gel electrophoresis 尿外泌体蛋白二维凝胶电泳增溶方法的比较

Journal of capillary electrophoresis Pub Date : 2012-01-01 DOI: 10.2198/JELECTROPH.56.7

Asami Kosaka, Akinobu Nakayama, Iku Yamaguchi, T. Kasama, M. Totsuka, K. Shiba

{"title":"Comparison of urinary exosomal protein solubilization methods for two-dimensional gel electrophoresis","authors":"Asami Kosaka, Akinobu Nakayama, Iku Yamaguchi, T. Kasama, M. Totsuka, K. Shiba","doi":"10.2198/JELECTROPH.56.7","DOIUrl":"https://doi.org/10.2198/JELECTROPH.56.7","url":null,"abstract":"Background: Urinary exosomal proteins have recently emerged as important candidates for elucidating the mechanisms underlying physiological events and disease-related metabolism in the kidney. Here, we evaluated standard sample preparation methods for two-dimensional gel electrophoresis (2DE) to determine which one yielded the maximum protein recovery from urinary exosomes for protein identification.Materials and Methods: Urinary exosomes were purified from a healthy subject by using ultracentrifugation. The final pellets were dissolved with PBS or RIPA buffer. After being desalted, these exosomal protein solutions were each treated with 1 of 4 rehydration buffers (Rbs) containing detergents in the following formulations: CHAPS (Rb1), CHAPS and Triton X-100 (Rb2), dodecyl maltoside (Rb3), and ASB-14 (Rb4).Results: For all Rbs, a much greater number of protein spots was detected in the samples isolated with RIPA than with PBS. Only minor differences were observed in the number of protein spots for Rb1-3. The largest protein spots were detected using the combination of RIPA buffer and Rb4; however, the background on the 2DE gel was high in the region of >66 kD and at the lower pH values. For all combinations, the co-precipitation of the urinary Tamm-Horsfall protein masked the protein spots in the 66-100 kD region.Conclusion: For extracting a large number of proteins with a relatively clear background on silver-stained 2DE gels, the optimal exosomal protein-dissolving buffer is RIPA buffer. All of the evaluated Rbs, except for the one containing ASB-14 as a detergent, is suitable for solubilizing exosomal proteins on 2DE.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"215 1","pages":"7-11"},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89338218","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

Development of Formulas for Estimation of Cholesterol Levels in Major Serum Lipoproteins Separated by Agarose Gel Electrophoresis 琼脂糖凝胶电泳分离的主要血清脂蛋白中胆固醇水平测定公式的建立

Journal of capillary electrophoresis Pub Date : 2011-01-01 DOI: 10.2198/JELECTROPH.55.23

S. Vanavanan, Sirirat Chaloeysup, K. Kotani, Pornpen Srisawasdi

{"title":"Development of Formulas for Estimation of Cholesterol Levels in Major Serum Lipoproteins Separated by Agarose Gel Electrophoresis","authors":"S. Vanavanan, Sirirat Chaloeysup, K. Kotani, Pornpen Srisawasdi","doi":"10.2198/JELECTROPH.55.23","DOIUrl":"https://doi.org/10.2198/JELECTROPH.55.23","url":null,"abstract":"Background/aim: Electrophoresis is useful for examining the lipoprotein fraction patterns. A simultaneous and cost-effective addition of cholesterol levels in each lipoprotein fraction to the lipoprotein patterns can more assist clinical decision-making. This study' aim was to develop the formulas for estimating the fractionated lipoproteins cholesterol levels in a recent system, the agarose gel Sebia HYDRAGEL LIPO+Lp(a) electrophoresis. Methods: Serum samples were analyzed by two Sebia electrophoresis, HYDRAGEL LIPO+Lp(a) and the quantitative HYDRAGEL LDL/HDL-CHOL Direct methods. The formulas for estimation of relative cholesterol (%) of individual lipoprotein fractions were developed using linear regression models. Thereafter, the calculated lipoproteins cholesterol values by multiplying the relative cholesterol with total cholesterol concentrations were compared with the standardized enzymatic assayed values. Results: The equations for calculating % relative cholesterol (y) from % relative lipoprotein (x) were y=x-8, x+21 and 0.75x-6.5 for high-density lipoprotein (HDL), low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL) fractions, respectively. Regression statistics obtained between the calculated assays (y) and the standardized enzymatic assays (x') in samples with and without Lp(a) were y=1.07x'- 0.18 and 1.06x'-0.06, respectively for HDL-cholesterol, y=0.90x'+0.32 and 0.92x'+0.29 for LDL-cholesterol, and y=0.85x'-0.03 and 0.95x'+0.02 for VLDL-cholesterol. Conclusions: The proposed formulas can provide a reliable estimation of cholesterol levels in each major lipoprotein fraction by the HYDRAGEL LIPO+Lp(a) electrophoresis. Further studies with its application are needed.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"36 3 1","pages":"23-29"},"PeriodicalIF":0.0,"publicationDate":"2011-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78472820","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

Variations of urinary protein excretion and α1-antitrypsin molecular size by glycemic control in type 2 diabetic patients 2型糖尿病患者尿蛋白排泄及α1-抗胰蛋白酶分子大小与血糖控制的关系

Journal of capillary electrophoresis Pub Date : 2011-01-01 DOI: 10.2198/JELECTROPH.55.13

Mayuka Goya, Mie Arai, K. Yamanaka, Y. Kanai, K. Shiba, Kenji Sato

{"title":"Variations of urinary protein excretion and α1-antitrypsin molecular size by glycemic control in type 2 diabetic patients","authors":"Mayuka Goya, Mie Arai, K. Yamanaka, Y. Kanai, K. Shiba, Kenji Sato","doi":"10.2198/JELECTROPH.55.13","DOIUrl":"https://doi.org/10.2198/JELECTROPH.55.13","url":null,"abstract":"We tried to determine the variations of levels and molecular heterogeneities of various proteins which occur in type 2 diabetes patients during short-term hospitalization for glycemic control. Type 2 diabetes patients were classified into 2 groups: only oral diabetes medicine therapy (group I) and only insulin and/or insulin plus oral diabetes medicine therapy (group II). All urinary proteins in group I tended to decrease on discharge day. Tamm-Horsfall protein (THP) levels of group II significantly increased on discharge day. There were significant correlations between each protein except with THP on day of admission and discharge of group I. In addition, there were no correlations between THP and other proteins except with α1-antitrypsin (α1-AT) in group II. Comparison of blood glucose level and HbA1c on admission day and first clinic day after discharge revealed that HbA1c level on discharge day was significantly lower in group II. We examined the molecular size of various proteins by SDS-PAGE and western blotting. In both groups, 9 α1-AT bands of different molecular sizes were detected besides a main band, but in group II, a 43 kDa α1-AT fragment, in urine collected closer to discharge day was found to be more densely stained than that in urine collected on admission day. These results revealed that glycemic control by insulin increased THP levels, remarkably reduced other protein levels, and caused urinary α1-AT to be of low molecular size.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"11 1","pages":"13-22"},"PeriodicalIF":0.0,"publicationDate":"2011-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83439651","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

Inverse-gradient polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate for better separation of protein samples 反梯度聚丙烯酰胺凝胶电泳在十二烷基硫酸钠存在下能更好的分离蛋白质样品

Journal of capillary electrophoresis Pub Date : 2011-01-01 DOI: 10.2198/JELECTROPH.55.1

M. Hashiguchi, K. Shimizu, T. Hashiguchi

引用次数: 1

Urinary protein profiles in patients with ureterolithiasis and nephrolithiasis 输尿管结石和肾结石患者的尿蛋白谱

Journal of capillary electrophoresis Pub Date : 2011-01-01 DOI: 10.2198/JELECTROPH.55.5

Yasuko Kawakami, Naoki Sakai, K. Shiba, Kenji Sato

{"title":"Urinary protein profiles in patients with ureterolithiasis and nephrolithiasis","authors":"Yasuko Kawakami, Naoki Sakai, K. Shiba, Kenji Sato","doi":"10.2198/JELECTROPH.55.5","DOIUrl":"https://doi.org/10.2198/JELECTROPH.55.5","url":null,"abstract":"In ureterolithiasis and nephrolithiasis patients, urinary total protein (TP), albumin (Alb), and IgG levels were significantly higher than those in control subjects, whereas THP was unchanged, Alb/TP ratio was significantly increased, and THP/TP ratio was significantly decreased in ureterolithiasis patients only. All proteins in patients with initial and recurrent ureterolithiasis were unchanged. However, TP and Alb levels in patients with recurrent nephrolithiasis were 10 times higher than those in patients with initial nephrolithiasis; IgG and THP levels were also higher in recurrent diagnosis patients. In patients with initial and recurrent ureterolithiasis, the rate of increase of urinary proteins before and after extracorporeal shockwave lithotripsy (ESWL) treatment was 8.9±12.5 and 8.4±1.7 fold. TP level of initial and recurrent nephrolithiasis patients before and after ESWL was 48.8±28.2 and 12.8±9.5 fold rate of increase, respectively. Alb level of initial patients was 58.9±50.4 fold rate of increase and that of recurrent patients was 9.9±8.2 fold. We observed changes in levels and molecular heterogeneity of Alb, THP, and IgG proteins. The Alb, THP, and IgG bands with various molecular sizes in patients were detected when compared to control subjects. This study suggested that the heterogeneity of Alb, THP, and IgG proteins may be involved in stone formation.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"19 1","pages":"5-12"},"PeriodicalIF":0.0,"publicationDate":"2011-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76192210","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

Expression of intestinal-type alkaline phosphatase mRNA in liver of Akp3 knockout mice Akp3基因敲除小鼠肝脏肠型碱性磷酸酶mRNA的表达

Journal of capillary electrophoresis Pub Date : 2010-01-01 DOI: 10.2198/JELECTROPH.54.27

M. Goseki‐Sone, N. Sogabe, Takanari Nakano, R. Tanabe, M. Haraikawa, D. Alpers, T. Komoda

{"title":"Expression of intestinal-type alkaline phosphatase mRNA in liver of Akp3 knockout mice","authors":"M. Goseki‐Sone, N. Sogabe, Takanari Nakano, R. Tanabe, M. Haraikawa, D. Alpers, T. Komoda","doi":"10.2198/JELECTROPH.54.27","DOIUrl":"https://doi.org/10.2198/JELECTROPH.54.27","url":null,"abstract":"Previously, we discovered that the rat liver showed enhanced expression of intestinal-type alkaline phosphatase (ALP) on high fat-feeding. Here we tested the hypothesis that intestinal-type ALP (Akp3 or Akp6) might also be regulated in the liver of Akp3-/- mice upon high fat-feeding. C57Bl/6J male Akp3-/-, Akp3+/-, or wild-type mice were fed a normal control or high-fat diet and the effects on intestinal-type ALP mRNA in liver were investigated. We found that Akp3 mRNA is only expressed in the intestine and not in the liver, while Akp6 mRNA is expressed in both the upper intestine and liver of wild-type mice. The intensity of Akp6 mRNA expression in the liver was not enhanced in the Akp3-/- mice compared with the wild-type mice fed on a high-fat diet. The nucleotide sequence of the PCR product of Akp6 from the liver was identified as being the same as that of Akp6 in the intestine. This is the first report concerning IAP mRNA expression in the mouse liver, but further studies will be needed to determine if this ectopic expression of intestinal-type ALP is associated with any unique function.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"75 1","pages":"27-32"},"PeriodicalIF":0.0,"publicationDate":"2010-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80809751","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

Dynamic analysis of the silver staining gel image of 2-DE for protein spots segmentation 2-DE银染色凝胶图像的动态分析，用于蛋白质斑点分割

Journal of capillary electrophoresis Pub Date : 2010-01-01 DOI: 10.2198/JELECTROPH.54.1

M. Kishimoto, Y. Tatsumi, N. Tamesui, Y. Kumada, J. Horiuchi, K. Okumura

引用次数: 3

A semiautomatic analyzer for urinary protein assay and a software program for classifying renal injury using cellulose acetate electrophoresis 一种半自动尿蛋白分析仪和一种醋酸纤维素电泳肾损伤分类软件程序

Journal of capillary electrophoresis Pub Date : 2010-01-01 DOI: 10.2198/JELECTROPH.54.13

R. Kubota, M. Ishii, M. Kameko, H. Kitamura, M. Sakatsume, Kenji Sato, K. Shiba

{"title":"A semiautomatic analyzer for urinary protein assay and a software program for classifying renal injury using cellulose acetate electrophoresis","authors":"R. Kubota, M. Ishii, M. Kameko, H. Kitamura, M. Sakatsume, Kenji Sato, K. Shiba","doi":"10.2198/JELECTROPH.54.13","DOIUrl":"https://doi.org/10.2198/JELECTROPH.54.13","url":null,"abstract":"The purpose of this study was to develop and evaluate a semiautomatic analyzer for total urinary protein assay and to develop a software program for identifying the affected part of the kidney by using data obtained by cellulose acetate electrophoresis (CAE) with silver staining. The limit of detection of this semiautomatic analyzer was 2.5 mg/L, and the assay precision was good, with a coefficient of variation (CV) less than 10%. Moreover, there was a good correlation between the results obtained by using this semiautomatic analyzer and the manual method. The urinary protein fractions of patients who were diagnosed with renal disease on the basis of renal biopsy were analyzed by performing CAE with silver staining. The relative mobility (RM) of several bands in the β fraction was calculated. The algorithm of the method used for calculating RM was programmed, and the RM ranges detected in several major protein bands were incorporated in the programmed software to classify the types of renal disorders. The programmed software could be used to classify the types of nephropathy in the case of diabetic patients with albumin concentrations of 30 mg/gCre or less. This semiautomatic analyzer can help detect nephropathy in early stages, and the urinary protein fraction analysis software can help determine the type of renal disorder in diabetic patients before microalbuminuria is detected, thus facilitating early treatment.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"37 1","pages":"13-18"},"PeriodicalIF":0.0,"publicationDate":"2010-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79158349","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: 3

Electrophoretic analysis of various urinary proteins in Japanese patients with Dent disease 日本登特病患者各种尿蛋白的电泳分析

Journal of capillary electrophoresis Pub Date : 2010-01-01 DOI: 10.2198/JELECTROPH.54.19

Kumiko Sawada, T. Matsuyama, T. Sekine, K. Shiba, Kenji Sato

{"title":"Electrophoretic analysis of various urinary proteins in Japanese patients with Dent disease","authors":"Kumiko Sawada, T. Matsuyama, T. Sekine, K. Shiba, Kenji Sato","doi":"10.2198/JELECTROPH.54.19","DOIUrl":"https://doi.org/10.2198/JELECTROPH.54.19","url":null,"abstract":"Patients with Dent disease excrete high amounts of urinary α1-microglobulin (α1-m), β2-microglobulin (β2-m), and retinol-binding protein (RBP). We found that the levels of α1-m, β2-m, and RBP in patients with Dent disease were higher than those in healthy subjects by 20 times, 130 times, and 200 times, respectively. These results confirmed the results of previous studies. The cellulose acetate membrane electrophoresis (CAE) showed that the characteristics of the urinary protein fraction in patients with Dent disease included appearance of pre-albumin, low albumin value, equal α1-globulin (α1-G) and α2-globulin (α2-G) values, disappearance of β-globulin (β-G), and appearance of slow α2-G (RBP) and slow β-G (β2-m). Urinary albumin (ALB), α1-m, β2-m, and RBP of patients with Dent disease 1 (mutation of CLCN 5) and Dent disease 2 (mutation of OCRL 1) and of healthy subjects were identified using western blot analysis after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Six bands from 84 kDa to 130 kDa were detected in addition to a main band of ALB (67 kDa). Seven bands from 51 kDa to 92 kDa were detected in addition to a main band of α1-m (34 kDa). A 17-kDa band was detected in addition to a main band of β2-m (14 kDa). RBP was detected only in the main band (17 kDa). The ALB and α1-m bands, which were in the high molecular weight range in addition to the main band, were densely stained in Dent disease 2.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"39 1","pages":"19-25"},"PeriodicalIF":0.0,"publicationDate":"2010-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86136258","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