bluuebunny/crossref_metadata_2025_split huggingface dataset:
The genetic haptoglobin polymorphism: Relevance of paternity assessment
Authors: Patzelt Dieter , Geserick Gunther , Schröder Helga 
Published: May 2005

AbstractA practical method for haptoglobin subtyping is described utilizing fast sample preparation by means of batch adsorption to DEAE‐cellulose and subsequent isoelectric focusing of reductively cleaved samples. The expanded haptoglobin polymorphism leads to an increase of the theoretical paternity exclusion rate to approximately 33 %. Hence, the system appears to be highly attractive for paternity assessment.
Analysis of the genetic polymorphism of coagulation factor XIIIB (FXIIIB) by isoelectric focusing
Authors: Leifheit Hans‐Jürgen , Cleve Hartwig 
Published: May 2005

AbstractThe genetic variants of the coagulation factor XIIIB (FXIIIB) were analyzed by isoelectric focusing, carried out in agarose gels and followed by immunofixation. The FXIIIB phenotypes were visualized by a combined staining procedure with Coomassie Brilliant Blue R‐250 and silver nitrate. Improved resolution was accomplished in polyacrylamide gels by hybrid isoelectric focusing in immobilized pH gradients supplemented with carrier ampholytes. We examined a total of 1.604 unrelated, healthy individuals from Southern Germany. The frequencies for the FXIIIB alleles were B*1 = 0.7581, B*2 = 0.0843, B*3 = 0.1568 and B*4 = 0.0019. The theoretical exclusion rate for disputed paternity is 22.35 %.
Two new esterase D(ESD) variants revealed by isoelectric focusing in agarose gel
Authors: Weidinger Sebastian , Henke Jürgen 
Published: May 2005

AbstractUsing isoelectric focusing in thin‐layer agarose gel (AGIF) with the narrow pH range of 4.5–5.4, a high resolution of esterase D (ESD) isozyme banding patterns has been achieved. Some variant phenotypes which could not be distinguished from common ESD types by conventional electrophoresis have shown different patterns after AGIF. The IEF method permitted the distinction of two further variants in the ESD system, tentatively named ESD Rehren and ESD Ravensburg. We recommend, therefore, that for the classification of ESD phenotypes a high resolution IEF technique should be used.
The use of α2HS‐glycoprotein and group specific component in typing forensic blood samples for discriminative and investigative purposes
Authors: Westwood Sara A. 
Published: May 2005

AbstractMethods are described for phenotyping α2HS‐glycoprotein (A2HS) and group specific component (GC) in plasma and blood‐stains. The methods have been developed to be sensitive, to provide unequivocal results and to give maximal information from minimal amounts of sample. In attempting to fulfill the last criterion, a new method is described for the simultaneous typing of α2 HS‐glycoprotein and group‐specific component from serum or plasma samples. The use of these proteins in determining the racial origin of a blood sample is also discussed.
The adsorption of large proteins in electrofocusing on immobilized pH gradients: II. Dependence on the oligomeric state of Immobiline
Authors: Fawcett John S. , Vicchio Domenick , Chrambach Andreas 
Published: May 2005

AbstractFive proteins with molecular mass in excess of 200 kDa were found to adsorb onto gels during isoelectric focusing on immobilized pH gradients (IPGEF). To probe for the mechanism of that adsorption, the homogeneity of the six Immobiline preparations used to make IPGEF gels was tested. Five of these Immobiline preparations appear homogeneous in gel filtration of Sephadex G‐10. The sixth Immobiline (pK 9.3) exhibits a minor component eluting ahead of the major peak and comprising less than 4 % of the total Immobiline absorbing at 226 nm. The proportion of the minor component increases with column load. Major and minor components when isolated appear to equilibrate with one another. Judging by the results of mass spectrometry, all 6 preparations are free of small aggregates of less than 500‐600 Da molecular mass. Ultrafiltration of the Immobiline preparations through a membrane with 500 Da nominal cutoff leads to partial desorption of only 3 of the 5 adsorbed proteins. CHAPS is ineffectual in desorbing the 5 proteins from the IPG gel made with ultrafiltered Immobilines. None of the 6 Immobiline preparations used precipitates ferritin. All large proteins that adsorb onto IPGEF gels in the pH range 4–9.5 also adsorb onto commercial IPGEF gels in the pH range 4–7.
Two‐dimensional electrophoresis. Dual‐label autoradiographic analysis of human skin fibroblast and myoblast proteins by two‐dimensional polyacrylamide gel electrophoresis using immobilised pH gradients in the first dimension
Authors: Patel Ketan , Dunn Michael J. , Günther Siegfried , Postel Wilhelm , Görg Angelika 
Published: May 2005

AbstractHorizontal two‐dimensional polyacrylamide gel electrophoresis with immobilised pH gradients in the first dimension has been applied to the analysis of human skin fibroblast and muscle myoblast total cell proteins. Excellent two‐dimensional separations of skin fibroblast proteins were obtained using pH 4–10 immobilised pH gradient gels with a long interelectrode distance (16 cm), but resolution was degraded, particularly of the more acidic proteins, by the use of shorter (10 cm) gels. Improved resolution of acidic and basic proteins was obtained using separate pH 4–7 and pH 7–10 immobilised pH gradient gels respectively in the first dimension. Two‐dimensional protein maps of skin fibroblast proteins were visualised both by silver staining and by autoradiography of samples labelled synthetically with [35S]methionine. Horizontal two‐dimensional electrophoresis, using pH 4–7 and pH 7–10 immobilised pH gradient gels in the first dimension, was applied to the analysis of protein samples from skin fibroblasts and muscle myoblasts dual‐labelled synthetically with [35S]methionine and [75Se]selenomethionine in an attempt to identify sets of proteins specific to each cell type. In addition, two‐dimensional maps or protein samples derived from normal individuals and patients with Duchenne muscular dystrophy were compared to search for protein changes associated with the disease state. Although sets of qualitative protein spot differences were observed by visual inspection of the two‐dimensional gels, more rigorous qualitative and quantitative analysis of the patterns using a computerised analysis system will be required to obtain the maximum amount of information from these data.
Preparation by two‐dimensional electrophoresis of proteins for antibody production: Antibodies against proteins whose synthesis is reduced by auxin in tobacco mesophyll protoplasts
Authors: Meyer Yves , Grosset Jean , Chartier Yvette , Cleyet‐Marel Jean‐Claude 
Published: May 2005

AbstractUsing phenol extraction from tobacco callus, we have prepared extracts with a high protein content. These proteins were separated in cylindrical non‐equilibrium pH gradient gels and visualized by dipping in sodium dodecyl sulfate (SDS)‐containing solution. Three gel sections, each containing proteins previously detected as abundantly synthesized in tobacco mesophyll protoplasts and whose synthesis is reduced by auxin application, were excised from each gel and collected. These proteins were further separated on slab SDS gels and protein bands were excised after Coomassie Brilliant Blue R‐250 staining and used to inject three rabbits. After one booster, highly specific antibodies were detected in their sera by ELISA and immunoblotting. Using these sera we have confirmed that the corresponding proteins are identical in callus and mesophyll protoplast and demonstrated that they are abundantly accumulated in tobacco roots but are undetectable in aerial organs and seeds.
Polypeptide patterns obtained by polyacrylamide gel electrophoresis of thylakoid membranes isolated from light and dark grown strains of Chlorogloea fritschii
Authors: Faulkner Michael 
Published: May 2005

AbstractSodium dodecyl sulphate‐polyacrylamide gel electrophoresis has been used to determine the difference between thylakoid polypeptide patterns of light and dark grown strains of the cyanobacterium Chlorogloea fritschii. There were only 2 prominent bands present in the dark grown strains, polypeptide Mr 50 000 and polypeptide Mr 90 000, also five fainter bands in the Mr range 45 000–66 200 corresponding to photosystem one, compared with the 32 bands present in the light grown strains. There was no obvious indication of the Mr 33 000 3‐(3,4‐chlorophenyl)‐1,1‐dimethyl‐urea binding protein. In addition the progressive daily development of the various photosystem components in the light and their relationship in photosynthesis was determined. It was observed that the increase of the relative concentrations of the photosystem two and phycocyanin components indicated their developments are mutually synchronized. The effect of light to dark and dark to light transfer on established strains was investigated. Appreciable loss of photosystem two components and the presence of an additional band Mr 22 500 of unknown function in the light to dark transfer, and little reactivation of the photosynthetic capabilities in the dark to light transfer was observed.
Report of workshop on cellular protein databases derived from two‐dimensional polyacrylamide gel electrophoresis
Authors: Neidhardt Frederick C. 
Published: May 2005

AbstractA workshop entitled Cellular Protein Databases from Two‐Dimensional Gel Electrophoresis was held in Atlanta, Georgia, 28 February–1 March 1987. Its purpose was to assess the status of two‐dimensional gel electrophoresis of proteins as a research methodology in biological systems, particularly in the generation of cellular protein databases. The workshop participants summarized current studies on a variety of biological systems, both prokaryotic and eukaryotic. Analysis of the progress being made led to the conclusion that electrophoretic techniques, supported by automatic scanning of gel images and computer‐assisted processing, analysis and matching of gel images, are now capable of generating databases of great potential value. Factors affecting the reproducibility of protein spot patterns on gels were identified, and the extent to which gel pattern variability causes difficulties in communicating results and in integrating information from different laboratories was assessed. Measures were suggested to help overcome obstacles to the generation of comprehensive cellular protein databases from the electrophoretic resolution of total cellular proteins.
Computerized, comprehensive databases of cellular and secreted proteins from normal human embryonic lung MRC‐5 fibroblasts: Identification of transformation and/or proliferation sensitive proteins
Authors: Celis Julio E. , Ratz Gitte P. , Madsen Peder , Gesser Borbala , Lauridsen Jette B. , Hansen Karl P. Brogaard , Kwee Sianette , Rasmussen Hanne Holm , Nielsen Henrik V. , Crüger Dorthe , Basse Bodil , Leffers Henrik , Honore Bent , Møller Olaf , Celis Ariana 
Published: May 2005

AbstractDatabases of protein information from human embryonal lung fibroblasts (MRC‐5) have been established using computer analyzed two‐dimensional gel electrophoresis. One thousand four hundred and eighty‐two cellular proteins (1060 with isoelectric focusing and 422 with nonequilibrium pH gradient electrophoresis, in the first dimension) ranging in molecular mass between 8 and 234 kDa were separated and numbered. Information entered in the database (in most cases for major proteins) includes: protein name, HeLa protein catalog number, mouse protein catalog number, proteins matched in transformed human epithelial amnion cells (AMA) and peripheral blood mononuclear cells (PBMC), transformation and/or proliferation sensitive proteins, synthesis in quiescent cells, cell cycle regulated proteins, mitochondrial and heat shock proteins, cytoskeletal proteins and proteins whose synthesis is affected by interferons. Additional information entered for a few transformation‐sensitive proteins that have been selected for future studies includes levels of synthesis and amounts in fetal human tissues. A total of four hundred and seventy‐six [35S] methionine labeled polypeptides (258 isoelectric focusing; 218, nonequilibrium pH gradient electrophoresis) secreted by MRC‐5 fibroblasts were separated and recorded (J. E. Celis et al., Leukemia 1987, 1, 707–717). Information entered in this database includes molecular weight and transformation sensitive proteins. These databases, as well as those of epithelial and lymphoid cell proteins (J. E. Celis et al., Leukemia 1988, 9, 561–601), represent the initial stages of a systematic effort to establish comprehensive databases of human protein information. In the long run, these databases are expected to offer a useful framework in which to focus the human genome sequencing effort.
Genomically linked cellular protein databases derived from two‐dimensional polyacrylamide gel electrophoresis
Authors: Neidhardt Frederick C. , Appleby David B. , Sankar Pushpam , Hutton M. Elizabeth , Phillips Teresa A. 
Published: May 2005

AbstractIn its most useful form a cellular protein database should be genomically based, because it is the genome which determines both the total number of proteins a cell can make and the particular ones that will be made under any given condition. Such a database should trace each protein back to its structural gene, and should account for every structural gene of a cell. Recent advances in molecular biology greatly facilitate the construction of such gene‐protein databases. The mapping of genes of unidentified proteins resolved from total cell extracts on two‐dimensional gels can now be accomplished by largely biochemical methods, without the necessity of isolating mutants or performing genetic crosses. Other techniques permit one to search gels for the product of any newly discovered gene (or open reading frame) suspected of encoding a protein. Consequently, gene‐protein indices can be built independently and simultaneously from either direction – deducing the genetic map from the protein pattern, or finding the protein pattern from information encoded in the genome. A database of this sort is being constructed for the bacterium, Escherichia coli. Given the current pace of DNA nucleotide sequencing, the development of total gene‐protein indices for a variety of cells can be anticipated in the near future.
A simple, sensitive method of analyzing bacterial ribosomal DNA polymorphism
Authors: Picard‐Pasquier Nicole , Ouagued Malika , Picard Bertrand , Goullet Philippe , Krishnamoorthy Rajagopal 
Published: May 2005

AbstractA rapid DNA extraction procedure and random primer labelling of Fscherichia coli ribosomal RNA with cloned reverse transcriptase have been used to establish a simple and highly sensitive method for studying ribosomal DNA polymorphism in bacteria. Examples of inter‐ and intraspecies differentiation of bacteria are given and potential applications in bacterial epidemiology and taxonomy are discussed.
Classification of mouse submaxillary gland esteroproteases by their substrate specificities
Authors: Kurosawa Nobuyuki , Ogita Zen‐Ichi 
Published: May 2005

AbstractThe submaxillary gland esteroproteases were separated by two‐dimensional polyacrylamide gel electrophoresis, and their súbstrate specificities were determined by histochemical staining procedures using cellulose acetate membranes. Twenty‐one proteolytically active enzymes were classified into four groups based on substrate specificities on benzoyl‐arginine‐ethylester (BAEE), N‐α‐benzoyl‐DL‐arginine‐p‐nitroanilide (BAPNA), and poly‐L‐lysine. These types were further divided into eight subgroups by their sensitivity to inhibitors and androgen dependence. These results suggest that eight groups of esteroproteases are present in the submaxillary gland of male mice.
Influence of a soluble anionic polymer on the electrophoresis of proteins
Authors: Shimura Kiyohito , Kasai Ken‐Ichi 
Published: May 2005

AbstractThe influence of a soluble anionic polymer on electrophoresis of proteins was studied in relation to the nonspecific ionic effect of an affinophore on application to affinophoresis. Zone electrophoresis of proteins was carried out in agarose gel in the presence of succinyl‐poly‐L‐lysine (degree of polymerization, 120) by using three electrophoresis buffers differing in ionic strength (0.06, 0.12 and 0.18) and pH (7.0 and 7.9). Proteins migrated as distinct single bands even in the presence of the polymer. The mobility of cationic proteins towards the cathode was first decreased and then increased towards the anode as the polymer concentration increased, while that of anionic proteins was not affected. The dependence of the apparent mobility changes of the proteins on the concentration of the polymer was treated quantitatively in the same way as affinity electrophoresis. The extent of the ionic interaction between a cationic protein and the polymer could be estimated as an apparent dissociation constant. It greatly depended on the ionic strength of the electrophoresis buffer. Except for the extremely cationic proteins such as lysozyme, the ionic interaction with up to 0.1 mM of the polymer could be practically suppressed by the use of 0.1 M sodium phosphate buffer (pH 7.0).
Enhanced immunodetection of blotted house dust mite protein allergens on nitrocellulose following blocking with tween 20
Authors: Tovey Euan R. , Ford Stephen A. , Baldo Brian A. 
Published: May 2005

AbstractThe effect of blocking nitrocellulose membranes with the nonionic detergent Tween 20 on the detection, by protein blotting, of IgE‐binding to house dust mite Dermatophagoides pteronyssinus allergens has been investigated. Tween blocking led to enhanced immunodetection of allergens despite removal of proteins from the membrane when compared to protein blocking agents which did not displace transferred components. The enhancement varied with the different mite components and, for one in particular, antigen Der p II, an increase of more than 100‐fold in IgE antibody binding occurred despite a concurrent loss of more than 90% of Der p II from the membrane. Both the enhancement of binding and loss of components from the membrane were dependent upon the time course of blocking and the concentration of Tween used.
The application of sodium dodecyl sulphate‐polyacrylamide gel electrophoresis to the taxonomic identification of the total body protein band profiles of Diplostomum spp. metacercariae (Digenea), parasites of fish eyes
Authors: Faulkner Michael 
Published: May 2005

AbstractSodium dodecyl sulphate‐polyacrylamide gel electrophoresis has been used to determine the metacercariae (Mc) total body protein band profiles of different Diplostomum spp. Four species of fish were investigated, roach (Rutilus rutilus) infected with D. spathaceum Mc, gywniad (Coregonus laveratus) infected with D. coregonus Mc, ruff (Gymnocephalus cernua) infected with a Diplostomum species related to D. gasterostei Mc, and perch (Perca fluviatilis) infected with D. gasterostei Mc. The four species of Diplostomum Mc were distinguished by three different bands of molecular weight, Mr 55 500, 53 500 and 52 000. A homology of polypeptide component distribution was evident for D. gasterostei Mc, from P. fluviatilis, and D. coregonus Mc, from C. laveratus, the latter showing reduced protein concentrations between bands Mr 32 000–40 000. Critical analysis of Mc polypeptide patterns showed no evidence of contamination with lens, retina or vitreous humour host eye protein. Gross morphological data for the parasites was also considered in relation to different band profiles obtained for the Diplostomum spp. Mc. Band profile analysis in conjunction with other taxonomic tests proved to be a useful tool in the identification of unknown species.
Analysis of porcine erythrocyte lysate proteins by horizontal two‐dimensional electrophoresis with immobilized pH gradient in the first dimension
Authors: Russ Ingolf , Günther Siegfried , Pirchner Franz 
Published: May 2005

AbstractHorizontal two‐dimensional gel electrophoresis with immobilized pH gradients in the first dimension resulted in excellent patterns of porcine erythrocyte lysate proteins. The overwhelming excess of hemoglobin was removed by chloroform‐ethanol extraction and the number of spots further increased by acetone precipitation, revealing considerably more protein spots than detected by analyzing the untreated sample. A total of 430 protein spots have been reproducibly obtained for further analysis of genetic variations. A number of enzyme systems have been identified by visual inspection and comparison with previous results. The existence of further multiple forms for other enzymes is also apparent.
Mobility surfaces for field‐inversion gel electrophoresis of linear DNA
Authors: Crater Glenn D. , Gregg Michael R. , Holzwarth G. 
Published: May 2005

AbstractThe mobility of linear DNA during field‐inversion gel electrophoresis was measured as a function of molecular weight Mr, pulse time t, and field strength E. Values of Mr between 48.5 and 194 kilobase pairs (kb), E from 5 to 14 V/cm and pulse times of 0.3 to 12 s were used. The data are presented as three‐dimensional surfaces of mobility: E:t for fixed Mr or graphs of mobility: Mr:t for fixed E. The surfaces are not smoothly increasing functions of E, Mr, or t but instead show a valley with minimum mobility and a steep rise in mobility as t increases. For a field of 10 V/cm, 1 % agarose gels, and 3:1 ratio of forward: back pulse time, the forward switching time t* at which the mobility changes most rapidly is given by t* =(0.034 ± 0.003) Mr for Mr in kb and t* in seconds. The data and equations delineate the best conditions to achieve a particular separation.
Sieving of double‐stranded DNA during agarose gel electrophoresis
Authors: Serwer Philip 
Published: May 2005

AbstractAgarose gel electrophoresis is used to fractionate linear, double‐stranded DNA by its length. Sieving of the gel is the cause of this fractionation and has been investigated by developing theoretical models and by quantifying sieving observed during electrophoresis. Here are reviewed the following aspects of the fractionation of linear, double‐stranded DNA by agarose gel electrophoresis: (1) the basic observations that qualitatively characterize these fractionations, (2) evidence for the deformation of DNA's random coil, (3) quantitative analysis of the relationship of observed electro‐phoretic mobility to the DNA's length, (4) theoretical models that have been developed to explain data presented in Sections 1–3, (5) observations not yet quantitatively explained by models, and (6) some aspects of the use of a variable electrical field (pulsed‐field gel electrophoresis) to improve separations.
Measurement of protein‐DNA interaction parameters by electrophoresis mobility shift assay
Authors: Fried Michael G. 
Published: May 2005

AbstractNative gel electrophoresis (mobility shift) assays may be used to obtain quantitative information about the site distribution, equilibria and kinetics of protein‐DNA interactions. These applications depend on the ability of the electrophoretic system to resolve the reaction components, and on their stabilities during the separation process Factors which affect the lifetimes and mobilities of protein‐DNA complexes during electrophoresis include reaction and electrophoresis buffer composition, pH, and ionic strength; the presence of low molecular weight effectors and enzymatic substrates; the nature and concentration of the gel matrix; the temperature; the molecular weights of protein and DNA; the stoichiometric ratios of their complexes; and the possibility of conformational and configurational isomerization of reaction components. We discuss how these factors influence the acquisition of quantitative data from electrophoretic patterns and band intensities, and present formulas for the estimation of equilibrium constants and rate constants for prototypical DNA‐protein interactions.