Journal of biochemical and biophysical methods最新文献_第10页

Interaction of loratadine with serum albumins studied by fluorescence quenching method 荧光猝灭法研究氯雷他定与血清白蛋白的相互作用

Journal of biochemical and biophysical methods Pub Date : 2007-08-01 DOI: 10.1016/j.jbbm.2007.04.001

Bo Zhou , Zu-De Qi , Qi Xiao , Jia-Xin Dong , Ye-Zhong Zhang , Yi Liu

引用次数: 74

Reduction of dehydroascorbic acid at low pH 低pH下脱氢抗坏血酸的还原

Journal of biochemical and biophysical methods Pub Date : 2007-08-01 DOI: 10.1016/j.jbbm.2007.04.007

Luka Wechtersbach, Blaž Cigić

{"title":"Reduction of dehydroascorbic acid at low pH","authors":"Luka Wechtersbach, Blaž Cigić","doi":"10.1016/j.jbbm.2007.04.007","DOIUrl":"10.1016/j.jbbm.2007.04.007","url":null,"abstract":"<div>Ascorbic acid and dehydroascorbic acid are unstable in aqueous solution in the presence of copper and iron ions, causing problems in the routine analysis of vitamin C. Their stability can be improved by lowering the pH below 2, preferably with metaphosphoric acid. Dehydroascorbic acid, an oxidised form of vitamin C, gives a relatively low response on the majority of chromatographic detectors, and is therefore routinely determined as the increase of ascorbic acid formed after reduction. The reduction step is routinely performed at a pH that is suboptimal for the stability of both forms. In this paper, the reduction of dehydroascorbic acid with tris-[2-carboxyethyl] phosphine (TCEP) at pH below 2 is evaluated. Dehydroascorbic acid is fully reduced with TCEP in metaphosphoric acid in less than 20 min, and yields of ascorbic acid are the same as at higher pH. TCEP and ascorbic acid formed by reduction, are more stable in metaphosphoric acid than in acetate or citrate buffers at pH 5, in the presence of redox active copper ions. The simple experimental procedure and low probability of artefacts are major benefits of this method, over those currently applied in a routine assay of vitamin C, performed on large number of samples.</div>","PeriodicalId":15257,"journal":{"name":"Journal of biochemical and biophysical methods","volume":"70 5","pages":"Pages 767-772"},"PeriodicalIF":0.0,"publicationDate":"2007-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jbbm.2007.04.007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26756057","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: 77

Possible utilization of in vitro synthesized mRNAs specifically expressed in certain tissues as standards for quantitative evaluation of the results of microarray analysis 可能利用体外合成的mrna在某些组织中特异性表达作为定量评价微阵列分析结果的标准

Journal of biochemical and biophysical methods Pub Date : 2007-08-01 DOI: 10.1016/j.jbbm.2007.04.004

Rei Kakuhata , Masahiro Watanabe , Takenori Yamamoto , Rie Akamine , Naoshi Yamazaki , Masatoshi Kataoka , Satoshi Fukuoka , Mitsuru Ishikawa , Toshihiko Ooie , Yoshinobu Baba , Tomoshige Hori , Yasuo Shinohara

{"title":"Possible utilization of in vitro synthesized mRNAs specifically expressed in certain tissues as standards for quantitative evaluation of the results of microarray analysis","authors":"Rei Kakuhata , Masahiro Watanabe , Takenori Yamamoto , Rie Akamine , Naoshi Yamazaki , Masatoshi Kataoka , Satoshi Fukuoka , Mitsuru Ishikawa , Toshihiko Ooie , Yoshinobu Baba , Tomoshige Hori , Yasuo Shinohara","doi":"10.1016/j.jbbm.2007.04.004","DOIUrl":"10.1016/j.jbbm.2007.04.004","url":null,"abstract":"<div>To examine the possible usefulness of in vitro synthesized RNA as standards in microarray analysis, we prepared full-length mRNAs encoded by 3 rat metabolic genes for heart/muscle type carnitine palmitoyltransferase I (M-CPTI), uncoupling protein (UCP1), and heart/muscle type fatty acid-binding protein (H-FABP). Artificial RNA samples were prepared by adding known amounts of these synthetic mRNAs to total RNA from rat liver, and transcript levels of various genes were compared between the prepared artificial RNA samples and total RNA samples of rat liver by using an Agilent oligo microarray system. Upon the addition of these synthetic RNAs, signals from the DNA spots corresponding to these 3 genes were elevated, but those from the DNA spots representing other genes were not markedly influenced. Using the ratio of the increase in signal intensity of DNA spot to the amount of added RNA, we estimated the expression levels of several genes and compared them with the absolute expression levels determined by calibrated Northern analysis. As a result, the absolute transcript levels of 3 genes encoding acidic ribosomal phosphoprotein P0, type-1 voltage-dependent anion channel (VDAC1), and type-2 glucose transporter (GLUT2) were successfully estimated by this procedure. Furthermore, genes specifically expressed in certain tissues such as UCP1 were concluded to be good candidates as standards for use in microarray analysis.</div>","PeriodicalId":15257,"journal":{"name":"Journal of biochemical and biophysical methods","volume":"70 5","pages":"Pages 755-760"},"PeriodicalIF":0.0,"publicationDate":"2007-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jbbm.2007.04.004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26731837","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: 12

A chromatographic tool for preparing combinatorial quinone–thiol conjugate libraries 一种制备组合醌-巯基偶联文库的色谱工具

Journal of biochemical and biophysical methods Pub Date : 2007-08-01 DOI: 10.1016/j.jbbm.2007.04.008

Maria Marti Villalba , Verity J. Litchfield , Robert B. Smith , Anthony M. Franklin , Callum Livingstone , James Davis

{"title":"A chromatographic tool for preparing combinatorial quinone–thiol conjugate libraries","authors":"Maria Marti Villalba , Verity J. Litchfield , Robert B. Smith , Anthony M. Franklin , Callum Livingstone , James Davis","doi":"10.1016/j.jbbm.2007.04.008","DOIUrl":"10.1016/j.jbbm.2007.04.008","url":null,"abstract":"<div>Quinones are well established as key players in the production of reactive oxygen species within cellular environments. Many factors govern their cytotoxicity but most studies have been restricted to a few, core, derivatives. A new strategy for the in situ production of quinone derivatives has been developed such that libraries of diverse functionality can be rapidly created without recourse to extensive synthetic procedures. The approach relies upon nucleophilic addition by reduced thiol derivatives to the quinone core within a pre-culture assay mixture and provides a generic strategy that exploits the large reservoir of commercial thiols currently available. A readily accessible chromatographic method has been developed that allows the derivatisation process to be easily monitored and the purity of the resulting one pot preparation to be assessed. The viability of the combinatorial approach has been fully validated through comparison with a range of quinone-S-conjugates prepared using conventional bench synthesis. The latter have been fully characterised.</div>","PeriodicalId":15257,"journal":{"name":"Journal of biochemical and biophysical methods","volume":"70 5","pages":"Pages 797-802"},"PeriodicalIF":0.0,"publicationDate":"2007-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jbbm.2007.04.008","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26799506","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: 6

Development of a novel nano-Invader DNA chip system 新型纳米入侵DNA芯片系统的研制

Journal of biochemical and biophysical methods Pub Date : 2007-08-01 DOI: 10.1016/j.jbbm.2007.05.012

Sachio Nomura , Masatoshi Kondo , Makoto Nagano , Kana Matsui , Toru Egashira

引用次数: 12

Correlative morphometric and electrochemical measurements of serotonin content in earthworm muscles 蚯蚓肌肉血清素含量的相关形态计量学和电化学测量

Journal of biochemical and biophysical methods Pub Date : 2007-08-01 DOI: 10.1016/j.jbbm.2007.02.007

Boglárka Takács , Mária Csoknya , Róbert Gábriel , Géza Nagy

{"title":"Correlative morphometric and electrochemical measurements of serotonin content in earthworm muscles","authors":"Boglárka Takács , Mária Csoknya , Róbert Gábriel , Géza Nagy","doi":"10.1016/j.jbbm.2007.02.007","DOIUrl":"10.1016/j.jbbm.2007.02.007","url":null,"abstract":"<div>Distribution of serotonin (5-HT) content of nervous fibers in both the somatic and the visceral muscle of Eisenia fetida have been investigated using immunocytochemical staining and voltammetric measurements. The somatic muscles in the body wall are richer innervated with serotoninergic fibers than the visceral ones in the pharynx and gizzard. The relative density of immunopositive fibers in the circular muscle layer of the body wall was found to be 2.73% while in the prostomium it was 1.02%. In the case of the muscle in pharynx 1.12% and in gizzard 1.28% density values were found. Differential Pulse Voltammetric (DPV) measurements with carbon fiber electrodes in the above mentioned muscle layers gave 272.5 nA, 135.0 nA, 122.5 nA, 137.5 nA peak heights, respectively. In the statistical analysis T-test was used at a confidence level of 95% (p  <!-->0.05). DPV current peak (ip) values reflect clearly the 5-HT concentration differences.Significant correlation was found between the innervation density and the ip values recorded in different areas. The ip values recorded at different times in different locations are determined by instantaneous serotonin concentration of the living tissue. As far as we know this is the first report using in vivo voltammetry investigating serotonin content in earthworm, E. fetida.</div>","PeriodicalId":15257,"journal":{"name":"Journal of biochemical and biophysical methods","volume":"70 5","pages":"Pages 713-720"},"PeriodicalIF":0.0,"publicationDate":"2007-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jbbm.2007.02.007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26728749","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

Fast and easy colorimetric tests for single mismatch recognition by PNA–DNA duplexes with the diethylthiadicarbocyanine dye and succinyl-β-cyclodextrin 二乙基硫二碳菁染料和琥珀酰-β-环糊精对dna - dna双链物单错配识别的快速简便比色试验

Journal of biochemical and biophysical methods Pub Date : 2007-08-01 DOI: 10.1016/j.jbbm.2007.04.003

Tullia Tedeschi , Stefano Sforza , Sheng Ye , Roberto Corradini , Arnaldo Dossena , Makoto Komiyama , Rosangela Marchelli

{"title":"Fast and easy colorimetric tests for single mismatch recognition by PNA–DNA duplexes with the diethylthiadicarbocyanine dye and succinyl-β-cyclodextrin","authors":"Tullia Tedeschi , Stefano Sforza , Sheng Ye , Roberto Corradini , Arnaldo Dossena , Makoto Komiyama , Rosangela Marchelli","doi":"10.1016/j.jbbm.2007.04.003","DOIUrl":"10.1016/j.jbbm.2007.04.003","url":null,"abstract":"<div>The 3,3′-diethylthiacarbocyanine (DiSC2(5)) dye is able to aggregate on full matched PNA–DNA duplexes by changing its absorption properties, which are manifested by an instantaneous colour shift from blue to purple. However the spontaneous aggregation of the dye also on mismatched duplexes and even on free PNA strands makes the test quite aspecific. Here it is demonstrated that the addition of succinyl-β-cyclodextrin (Succ-β-CyD) to the solutions containing PNA–DNA duplexes and the dye strongly enhances the specificity of the colour shift, allowing for a fast, very specific and extremely sensitive visual recognition of mismatches in DNA strands by using PNA probes in combination with the DiSC2(5) dye. The phenomenon has been studied by CD and NMR spectroscopies. The method has been optimized and preliminarily applied for the recognition of an apoE gene mutation in human DNA samples.</div>","PeriodicalId":15257,"journal":{"name":"Journal of biochemical and biophysical methods","volume":"70 5","pages":"Pages 735-741"},"PeriodicalIF":0.0,"publicationDate":"2007-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jbbm.2007.04.003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26741706","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: 6

A high pressure liquid chromatography-based assay for glutathione-S-transferase class distinction assay 基于高压液相色谱的谷胱甘肽- s -转移酶分类分析

Journal of biochemical and biophysical methods Pub Date : 2007-08-01 DOI: 10.1016/j.jbbm.2007.04.005

Brian N. Blanchette, Bal Ram Singh

{"title":"A high pressure liquid chromatography-based assay for glutathione-S-transferase class distinction assay","authors":"Brian N. Blanchette, Bal Ram Singh","doi":"10.1016/j.jbbm.2007.04.005","DOIUrl":"10.1016/j.jbbm.2007.04.005","url":null,"abstract":"<div>In order to expedite the process of classification of the members of the family of glutathione-S-transferases (GSTs) high performance liquid chromatography with photodiode array detection (HPLC-PDA) was used as a means for measuring enzymatic activity. The GST chosen for the development of the HPLC-PDA technique was from equine liver (E-GST). The characterizing substrates, ethacrynic acid (EA) and bromosulfophthalein (BSP), along with previously gathered characterization data allowed for the distinction of α, μ or π-class enzymes. In an initial characterization of the previously unclassified E-GST it was determined that the enzyme was of the π-class with specific activities of 0.062, ±0.0015 μmol min−1 mg−1 and 0.0019, ±0.00064 μmol min−1 mg−1 for EA and BSP, respectively. Finally, the activity of the E-GST with the EA and BSP substrates, was measured by HPLC-PDA, and was found to be 0.027, ±0.003 μmol min−1 mg−1 and 0.002, ±0.0005 μmol min−1 mg−1, respectively. While the HPLC-PDA data do not mirror the spectrophotometric results quantitatively the overall response by the E-GST was the same. In general, the E-GSTs were shown to belong to the π-class when characterized by HPLC-PDA due to an EA specific activity greater than 0.01 μmol min−1 mg−1 and a negligible BSP activity (≤0.002 μmol min−1 mg−1).</div>","PeriodicalId":15257,"journal":{"name":"Journal of biochemical and biophysical methods","volume":"70 5","pages":"Pages 761-765"},"PeriodicalIF":0.0,"publicationDate":"2007-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jbbm.2007.04.005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26752545","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

Comparison of five methods for determination of total plasma protein concentration 五种测定血浆总蛋白浓度方法的比较

Journal of biochemical and biophysical methods Pub Date : 2007-08-01 DOI: 10.1016/j.jbbm.2007.05.009

Burcu Okutucu, Ayşşe Dınçer, Ömer Habib, Figen Zıhnıoglu

{"title":"Comparison of five methods for determination of total plasma protein concentration","authors":"Burcu Okutucu, Ayşşe Dınçer, Ömer Habib, Figen Zıhnıoglu","doi":"10.1016/j.jbbm.2007.05.009","DOIUrl":"10.1016/j.jbbm.2007.05.009","url":null,"abstract":"<div>Quantitation of exact total protein content is often a key step and is common to many applications in general biochemistry research and routine clinical laboratory practice. Before embarking on any type of protein analysis, particularly comparative techniques, it is important to accurately quantitate the amount of protein in the sample. In order to assess the quality of total protein estimation results, five methods were tested and were applied to the same pooled plasma sample. For this aim, Bradford (Coomassie Brilliant Blue), Lowry (Folin–Ciocalteau), Biüret, Pesce and Strande (Ponceau–S/TCA), and modified method of Schaffner–Weismann (Amido Black 10B) were used. The last two methods employ simultaneous precipitation of proteins with the acid containing dye solutions followed by dissolution of precipitate in a NaOH solution. It is shown that each assay has advantages and disadvantages relative to sensitivity, ease of performance, acceptance in literature, accuracy and reproducibility/coefficient of variation. All of the methods tested show a CV %  6. Besides pooled plasma, a known concentration of human serum albumin was also analyzed and discussed by means of standardization of plasma total protein content.</div>","PeriodicalId":15257,"journal":{"name":"Journal of biochemical and biophysical methods","volume":"70 5","pages":"Pages 709-711"},"PeriodicalIF":0.0,"publicationDate":"2007-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jbbm.2007.05.009","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26799507","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: 139

Generic method for quantification of FLAG-tagged fusion proteins by a real time biosensor 用实时生物传感器定量flag标记融合蛋白的通用方法

Journal of biochemical and biophysical methods Pub Date : 2007-06-10 DOI: 10.1016/j.jbbm.2007.01.004

Christa Mersich, Alois Jungbauer

引用次数: 10