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Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society最新文献

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The biased reptation model of DNA gel electrophoresis: a user guide for constant field mobilities. DNA凝胶电泳的有偏重复模型:恒定场移动的用户指南。
Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01
G W Slater, P Mayer, S J Hubert, G Drouin
{"title":"The biased reptation model of DNA gel electrophoresis: a user guide for constant field mobilities.","authors":"G W Slater,&nbsp;P Mayer,&nbsp;S J Hubert,&nbsp;G Drouin","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The biased reptation model of DNA gel electrophoresis is simple enough to allow one to obtain detailed analytical and numerical predictions for experimentally relevant situations. Although it is not always applicable for explaining experimental results, the biased reptation model is usually a good starting point for data analysis. Unfortunately, the model is often reported as being incapable of explaining experimental data because the users have not analyzed the data properly or because they attempted to use the model outside its expected range of applicability. This article presents a detailed practical guide to the model and its limitations, as well as a complete description of its predictions regarding the analysis of constant field mobilities.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 2","pages":"71-9"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18877686","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
High resolution 2-D peptide mapping with subsequent analysis of peptides by microsequencing or lectin binding directly from PVDF membrane blots. 通过微测序或直接从PVDF膜印迹进行凝集素结合的高分辨率二维肽图谱分析。
Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01
M F Lopez, P Barry, W B Sawlivich, T Hines, W M Skea
{"title":"High resolution 2-D peptide mapping with subsequent analysis of peptides by microsequencing or lectin binding directly from PVDF membrane blots.","authors":"M F Lopez,&nbsp;P Barry,&nbsp;W B Sawlivich,&nbsp;T Hines,&nbsp;W M Skea","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Peptide mapping is a technique that is frequently used to characterize proteins. Typically, the method involves the cleavage of proteins in solution or in a gel with the subsequent separation of the peptide fragments on a 1-D SDS PAGE gel. Electrophoretic peptide maps are often used to compare homologous proteins from related organisms to derive evolutionary relationships. Other applications of peptide mapping include immunoblotting studies of selected proteins. Two-dimensional peptide mapping, a less common technique, has traditionally involved a combination of thin layer or paper chromatography and electrophoresis. Amino acid sequencing of peptides that were separated using this method and then subsequently blotted to PVDF membrane was reported recently. However, the resolution achieved with these methods is far below that which can be achieved with conventional 2-D electrophoresis of proteins in polyacrylamide gels. This report describes an electrophoretic system for the high resolution 2-D separation of peptides in gels with subsequent blotting to a novel cationic PVDF membrane, Immobilon-CD, and microsequencing directly from the 2-D blot. In addition, the high resolution peptide maps can be further analyzed by techniques such as lectin probing to determine post-translational modifications.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 2","pages":"95-102"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18877689","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
Two-dimensional electrophoretic study of normal colon mucosa and colorectal cancer. 正常结肠黏膜和结直肠癌的二维电泳研究。
Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01
H Kovarova, J Stulík, D F Hochstrasser, J Bures, B Melichar, P Jandík
{"title":"Two-dimensional electrophoretic study of normal colon mucosa and colorectal cancer.","authors":"H Kovarova,&nbsp;J Stulík,&nbsp;D F Hochstrasser,&nbsp;J Bures,&nbsp;B Melichar,&nbsp;P Jandík","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Two-dimensional gel electrophoresis has been used to analyze polypeptide composition of specimens of human normal colon mucosa and colorectal carcinomas. Immobilized pH gradient in the first dimension improved the separation and enabled us to analyze proteins from whole tissue samples with high reproducibility. Polypeptides were separated within sigmoidal 3.5-10 pH gradient and range of mol. weight 15-200 KD. In global, protein patterns of normal and malignant tissue seemed to be close to each other. We attempted to combine two-dimensional electrophoretic separation with the immunoblotting technique using patient sera to identify possible tumor antigens. From a wealth of spots on silver stained protein maps of adjacent normal colon mucosa or tumor tissue, only few spots gave positive reaction. These spots might be classified into two groups: 1) common for normal colon mucosa and tumor, 2) specific for tumor tissue. The extent of patient's antibody reaction with antigens appeared to correspond to urinary neopterin level, an index of immune activation.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 3","pages":"103-6"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18619476","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
On the "door-corridor" model of gel electrophoresis. II. Developments related to new gels, capillary gel electrophoresis and gel chromatography. 关于凝胶电泳的“门-走廊”模型。2新型凝胶、毛细管凝胶电泳和凝胶色谱的研究进展。
Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01
B Kozulić
{"title":"On the \"door-corridor\" model of gel electrophoresis. II. Developments related to new gels, capillary gel electrophoresis and gel chromatography.","authors":"B Kozulić","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>An excellent separation of small DNA fragments is possible also at a low total polymer concentration. This is illustrated with novel gels containing 1% of the cross-linked agarose polymers. In gel-filled capillaries, preelectrophoresis in the direction opposite to that of separation may create gel discontinuities capable not only of separating, but also of concentrating the migrating molecules. Such capillaries are able to provide an efficiency in the range of 100 x 10(6) theoretical plates per meter. It is proposed that electrophoretic separations in gel-filled capillaries have to be performed at high field strengths in order to achieve the optimal ratio between gel resistance and electrokinetic forces. When considered in terms of the forces exerted on the gel polymers by the migrating macromolecules, gel filtration and gel electrophoresis appear as two profoundly different techniques.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 3","pages":"137-48"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18619480","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 snake venoms by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and two-dimensional electrophoresis. 十二烷基硫酸钠-聚丙烯酰胺凝胶电泳及双向电泳分析蛇毒。
Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01
T Marshall, K M Williams
{"title":"Analysis of snake venoms by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and two-dimensional electrophoresis.","authors":"T Marshall,&nbsp;K M Williams","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The protein composition of snake venom has been analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and high resolution two-dimensional electrophoresis (2-DE) with comparison of our simplified Tris/glycine/SDS (TGS) method with a commercial Tris/Tricine/SDS (TTS) gel system. SDS-PAGE suggests differences between species and similarities between related species but 2-DE indicates that the protein composition of the venoms is highly complex. The TTS system improves resolution upon 2-DE but our simplified TGS method is optimal for silver staining which, contrary to previous reports, enhances detection 100-fold.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 1","pages":"25-31"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18810971","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
Poly(ADP-ribose) polymerase in HeLa cells--a high resolution two-dimensional gel analysis. HeLa细胞中的聚(adp -核糖)聚合酶-高分辨率二维凝胶分析。
Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01
S Prasad, V Notario, A Dritschilo
{"title":"Poly(ADP-ribose) polymerase in HeLa cells--a high resolution two-dimensional gel analysis.","authors":"S Prasad,&nbsp;V Notario,&nbsp;A Dritschilo","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>PADPRP is an eukaryotic enzyme responsible for poly(ADP-ribosyl)ation of several chromatin-associated proteins. Consequently, the modified proteins acquire various lengths of negatively charged, covalently bound oligo or poly(ADP-ribosyl) homopolymers. The present study was undertaken to get an insight into the charge and size heterogeneity of the various auto-modified species of PADPRP and other protein acceptors of (ADP-ribose) polymers. Toward this end, we analyzed HeLa cells using high resolution two-dimensional gel electrophoresis (2D-PAGE). Resolution of HeLa total cellular lysates in the basic pH range combined with immunoblots and activity-blots revealed extensive modification and processing of the enzyme. In addition to the native enzyme, a protein of approximately 116 kDa, we observed several proteins that exhibit immunoreactivity to an antibody prepared against a peptide encompassing the N-terminal 20 amino acids of the polymerase. Several protein species showed auto-modifying potential in an in situ activity blot assay. We have also localized the ADP-ribosylated proteins in permeabilized HeLa cells to demonstrate protein acceptors of poly(ADP-ribose).</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 1","pages":"3-10"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18810972","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 biomolecules by electrophoretic analysis of reversible interactions. 通过可逆相互作用的电泳分析来表征生物分子。
Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01
N H Heegaard
{"title":"Characterization of biomolecules by electrophoretic analysis of reversible interactions.","authors":"N H Heegaard","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 2","pages":"43-63"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18877684","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
Charge modification in rodent hepatic Grp78/BiP following exposure to structurally diverse peroxisome proliferators. 暴露于结构不同的过氧化物酶体增殖体后啮齿动物肝脏Grp78/BiP的电荷改变。
Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01
F A Witzmann, B M Jarnot, J W Clack
{"title":"Charge modification in rodent hepatic Grp78/BiP following exposure to structurally diverse peroxisome proliferators.","authors":"F A Witzmann,&nbsp;B M Jarnot,&nbsp;J W Clack","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>This investigation was conducted to determine the comparative effect of structurally diverse peroxisome proliferators (PP) on the two-dimensional protein pattern of rat liver whole homogenates. Perfluoro-n-decanoic acid (PFDA), perfluoro-n-octanoic acid (PFOA), clofibrate, and di(2-ethylhexyl)phthalate(DEHP) are all known to cause the proliferation of hepatic peroxisomes and the induction of peroxisomal beta-oxidative and microsomal omega-oxidative enzymes. To detect potential differences between these compounds with regard to the liver, we examined the unique patterns of protein alteration produced by in vivo exposure to them. Following exposure to various doses, whole liver homogenates were prepared and separated by two-dimensional gel electrophoresis (2DE) using the ISO-DALT System. Stained gels were digitized and protein patterns analyzed using the Kepler 2D Gel Analysis System. Immunoglobulin heavy-chain binding protein (BiP), also known as 78 kD glucose regulated protein (Grp78), was identified immunologically and by comigration of recombinant Grp78. BiP is a luminal endoplasmic reticular (ER) protein that functions in the assembly and folding of nascent proteins as they enter the ER. The present results suggest a selective posttranslational modification of BiP following PFDA exposure. Single-dose exposure to PFDA was associated with a notable charge-modification of BiP that persists up to 30 days. PFOA, clofibrate, and DEHP had less effect in this regard. Our data suggest the likely nature of this PFDA-associated protein modification is associated with protein-phosphorylation. These results document the unique nature of PFDA's hepatotoxicity with respect to classic peroxisome proliferators and support the utility of 2D gel analysis in toxicity testing.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 2","pages":"81-8"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18877687","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
The biased reptation model of DNA gel electrophoresis: a user guide for constant field mobilities. DNA凝胶电泳的有偏重复模型:恒定场移动的用户指南。
Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01 DOI: 10.1007/978-1-4613-0543-9_41
G. Slater, P. Mayer, S. Hubert, G. Drouin
{"title":"The biased reptation model of DNA gel electrophoresis: a user guide for constant field mobilities.","authors":"G. Slater, P. Mayer, S. Hubert, G. Drouin","doi":"10.1007/978-1-4613-0543-9_41","DOIUrl":"https://doi.org/10.1007/978-1-4613-0543-9_41","url":null,"abstract":"","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"77 12","pages":"71-9"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50962822","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
Electrically controlled focusing of proteins and ampholytes between two modified electrolytes. Computer simulation. 两种改性电解质之间的蛋白质和两性电解质的电控聚焦。计算机模拟。
Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01
M Deml, J Pospíchal
{"title":"Electrically controlled focusing of proteins and ampholytes between two modified electrolytes. Computer simulation.","authors":"M Deml,&nbsp;J Pospíchal","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Properties of the focusing in simple electrolyte systems using four-pole electrophoretic capillary column was evaluated by a computer simulation. Simulated concentration, pH and conductivity profiles confirm the previous experimentally observed features of the technique, i.e., high concentration capabilities, separation of the ampholytes and proteins into individual adjacent zones, sharp profile of zones and presence of only background electrolyte in the zone. Moreover, the influence of the magnitude of the solvolytic fluxes on the position of the zones in the column is demonstrated.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 3","pages":"107-15"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18619477","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
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