Electrophoresis in 2004: The Annual Meeting of the American Electrophoresis Society

Protein phosphorylation plays a critical role in the signaling pathways regulating water and solute transport in the distal renal tubule (i.e., renal collecting duct). A central mediator in this process is the antidiuretic peptide hormone arginine vasopressin, which regulates a number of transport proteins including water channel aquaporin-2 and urea transporters (UT-A1 and UT-A3). Within the past few years, tandem mass spectrometry-based proteomics has played a pivotal role in revealing global changes in the phosphoproteome in response to vasopressin signaling in the renal collecting duct. This type of large-scale ‘shotgun’ approach has resulted in an exponential increase in the number of phosphoproteins known to be regulated by vasopressin and has expanded on the established signaling mechanisms and kinase pathways regulating collecting duct physiology. This article will provide a brief background on vasopressin action, will highlight a number of recent quantitative phosphoproteomic studies in both native rat kidney and cultured collecting duct cells, and will conclude with a perspective focused on emerging trends in the field of phosphoproteomics.     

电泳梳的改进

用这种改进的电泳梳做垂直板凝胶电泳更方便、更精确。     

基于低成本聚甲基丙烯酸甲酯(PMMA)电泳芯片的微型DNA分析仪

在低成本的聚甲基丙烯酸甲酯(PMMA)电泳芯片上,利用双通道共聚焦激光诱导荧光检测系统实现了单链DNA快速高效的分离检测．选用0.8 mm厚度的PMMA薄片加工微管道,一方面降低了检测限,另一方面提高了散热性能．通过线性聚丙烯酰胺筛分胶液以及使用纤维素衍生物对微管道表面进行动态修饰等条件的优化,芯片完成了高分辨率、高重现性的短串联重复序列(STR)等位基因的快速分型检测．两个STR位点D13S317 和CSF1PO的等位基因分型标准物(allelic ladder)和实际样本的PCR扩增产物均在3 min内达到了基线分离,表明低成本的PMMA电泳芯片在法医学及临床医学领域的基因分析方面具有良好的发展潜能．     

Preparative Cell Electrophoresis

Abstract

A simple method is described for preparative cell electrophoresis. Solid glass beads (? 0.1 mm diameter) are used as packing material in a cylindrical glass column, the interstices of which contain a citric acid-phosphate buffer of pH 7.6 and 0.015 ionic strength. Cell mixtures containing ? 6 × 10⁹ erythrocytes in 0.2 ml are electrophoresed at 25 V/cm for 1–2 hours after which the column contents are extruded and the separated cell fractions recovered. Complete separation was achieved between mixtures of human and chicken erythrocytes and untreated and papain-treated human erythrocytes. The electrophoretic mobilities of cells subjected to this procedure are in good agreement with previously published mobilities, obtained by microelectrophoresis.     

Biospecific Interaction in Electrophoresis

Aspects of practical assessment such as question design and management of the assessment are discussed. Reference is made to some of the problems and to the possibilities for innovation which are possible as a result of implementing practical assessment of the kind required by the GCSE examining groups.     

Electrophoresis using ultra-high voltages

Optimization of electrophoretic techniques is becoming an increasingly important area of research as microdevices are now routinely adapted for numerous biology and engineering applications. The present work seeks to optimize electrophoresis within microdevices by utilizing ultra-high voltages to increase sample concentration prior to separation. By imaging fluorescently-tagged DNA samples, the effects of both conventional and atypical voltage protocols on DNA migration and separation are readily observed. Experiments illustrate that short periods of high voltage during electrophoretic injection do not destroy the quality of DNA separations, and in fact can enhance sample concentration five-fold. This study presents data that illustrate increases in average resolution, and resolution of longer fragments, obtained from electrophoretic injections utilizing voltages between 85 and 850 V/cm.     

毛细管无胶筛分电泳

围绕着毛细管无胶筛分电泳的介质和机理,概括介绍了近年来这种技术在各个方面的发展,及其在DNA片段的分离、PCR扩增产物的检测和蛋白质分子量的测定等方面的应用前景．     

Electrophoresis of algal glucans

The ability of reactive dyes based on chloro-s-triazine to reactwith the primary and secondary hydroxyl groups of glucose, formingcovalent bonds, has been used to dye the storage polyglucosidesof the algae, Cytmidium caldarium, Oscillatoria princeps, Rhodymeniapertusa and Spirogyra setiformis, prior to electrophoresis oncellulose polyacetate strips in borate buffer. The techniqueresolves mixtures of glucans and defines some of the parametersof the internal structure of these glucans. The similarity ofthe glucans from Cyanidium caldarium and Oscillatoria princepsis evident using this method. The two components of the "starch"from Spirogyra setiformis have been studied and identified bytheir reactions with the dye and the mobilities of the dyedcomponents during electrophoresis. Inferences as to the biphyleticevolutionary pathways leading from the Cyanophyceae to the Rhodophyceaeand the Chlorophyceae can be made based on the data. (Received June 27, 1973; )     

Electrophoresis of adsorbed DNA

The electrophoresis mobilities of native calf thymus DNA adsorbed on the charged solid particles were measured by a micro-electrophoretic method as functions of pII, ionic strength, and DNA concentration. The mobility data confirm the adsorption of DNA both on the positively charged alumina and negatively charged resin particles at wide range of pH and ionic strength. The mobility data also indicate significant DNA adsorption by negatively charged glass in the acidic range of pH. The electrophoretic mobilities of DNA adsorbed on different substrate particles under identical conditions do not differ widely, indicating the major role of the adsorbed DNA rather than the covered substrate in controlling the charge behavior of the particle. The mobilities of the adsorbed DNA at salt pH are of a comparable order of magnitude to those for the dissolved DNA in solution. The mobility of the adsorbed heat-denatured and alkali-denatured DNA is lower than that of the native adsorbed DNA under identical conditions of pH and ionic strength.     

Electrophoresis of isolated mitochondria

Isolated respiring mitochondria suspended in an isotonic medium migrate to the anode when they are exposed to an electric field. A simple method has been devised to visualize this electrical property of the organelles utilizing a modified zone electrophoresis technique. Uncouplers of oxidative phosphorylation inhibit this anaphoretic property of isolated mitochondria.