bluuebunny/crossref_metadata_2025_split huggingface dataset:
A quantitative theory of the stern electric double layer
Authors: Michov Budin M. 
Published: May 2005

AbstractA quantitative theory of the Stern electric double layer is suggested. It is based on the view that every ion possesses a geometrical and an electrokinetic radius, that the ionic atmosphere begins from the geometrical one, and that the difference between these radii is the Stern quantity Δ. The equations of the mentioned radii and the quantity Δ are established and the values of the different potentials characterizing an ion and its ionic atmosphere are determined.
Enhancing the sensitivity of DNA detection and recovery from agarose gels
Authors: Georges Fawzy , Girard Sophie , Chibbar Ravindra N. , Constabel Friedrich 
Published: May 2005

AbstractBy inserting nitrocellulose strips into agarose gels alongside the electrophoresed lanes and passing an electric current perpendicularly in the direction of the strips, highly efficient transfer of DNA bands onto the membrane in the form of concentrated dots is achieved. DNA detection limits by this technique are enhanced, at least three times as visualized by ethidium bromide fluorescence and at least twice more by radiolabeling.
A motor‐driven syringe‐type gradient maker for forming immobilized pH gradient gels
Authors: Fawcett John S. , Sullivan James V. , Chidakel Burt E. , Chrambach Andreas 
Published: May 2005

AbstractA motor driven gradient maker based on the commercial model (Jule Inc., Trumbull, CT) was designed for immobilized pH gradient gels to provide small volumes, rapid stirring and delivery, strict volume and temperature control and air exclusion. The device was constructed and by a convenient procedure yields highly reproducible gradients either in solution or on polyacrylamide gels.
Improved transfer of high molecular weight proteins using fabric‐reinforced soft polyacrylamide gels
Authors: Starita‐Geribaldi Mireille , Giudicelli Jean , Sudaka Pierre 
Published: May 2005

AbstractRapid transfer of electrophoretically separated, high molecular weight proteins from fabric‐reinforced, soft, low‐concentration polyacrylamide gels (3.5 %T, 2.6 %C) is described. Polyester fabrics proved useful for gel reinforcement during vertical slab gel electrophoresis. The fabric‐reinforced gel retains its dimension during transfer, equilibration and drying, facilitating comparison of protein bands in the separation gel and transfer matrix.
Application of Neuhoff's optimized Coomassie Brilliant Blue G‐250/ammonium sulfate/phosphoric acid protein staining to ultrathin polyacrylamide gels on polyester films
Authors: Peisker Klaus 
Published: May 2005

AbstractAn optimized Coomassie staining procedure, utilizing Coomassie Brilliant Blue G‐250 in phosphoric acid/ammonium sulfate, was applied to ultrathin‐layer isoelectric focusing in 0.18 mm polyacrylamide gels, and sodium dodecyl sulfatepolyacrylamide gel electrophoresis in 0.38 mm polyacrylamide gels, both backed to Gel‐Fix polyester supporting films. After isoelectric focusing staining of gelatin and acidic proteins was better with the phosphoric acid/ammonium sulfate procedure than with conventional organic solvent methods. When applied to gels after sodium dodecyl sulfate‐polyacrylamide gel electrophoresis the sensitivity of the phosphoric acid/ammonium sulfate method was equal to that on conventional staining but lower than on silver staining.
Linear Ferguson plots of polystyrene sulfate size standards for the quantitative agarose gel electrophoresis of subcellular particles
Authors: Hahn Elisabeth , Wurts Linda , Tietz Dietmar , Chrambach Andreas 
Published: May 2005

AbstractAccurately standardized commercial polystyrene sulfate particles in agarose gel electrophoresis yield linear Ferguson plots at pH 7.4 over a gel concentration range up to 0.9% agarose which do not exhibit any significant sigmoidal curve elements, using either a discontinuous buffer system or a continuous buffer. Ferguson plots of these standard‐sized particles were evaluated using alternatively a linear or convex model, by means of a newly developed set of programs (to be used in conjunction with program M‐LAB) which (i) is sufficiently user‐friendly to allow for quantitative agarose gel electrophoresis of subcellular‐sized spherical particles based on their convex Ferguson plots with the same operational simplicity previously available for linear Ferguson plots only; (ii) simultaneously and interactively analyzes the Ferguson plots of all particles under consideration on the basis of an extended Ogston model.
Comparison between isoelectric focusing methods for the detection of orosomucoid phenotypes
Authors: Montiel Maria Dolores , Carracedo Angel , Lopez‐Rodriguez Isidro , Rodriguez‐Calvo Maria Sol , Concheiro Luis , Huguet Emilio , Gené Manuel 
Published: May 2005

AbstractOrosomucoid (ORM) polymorphism was investigated by different methods including isoelectric focusing in acid pH ranges followed by silver staining, print immunofixation of desialyzed ORM, fixation using a lectin from the sea‐weed Codium tomentosum, isoelectric focusing followed by immunofixation in miniaturized gels and isoelectric focusing in immobilized pH gradients. Population genetics studies were carried out in Galicia (NW Spain) and two new ORM variants were found.
Detection of turnip crinkle virus on agarose gel electropherograms at the nanogram load level
Authors: Orbán László , Chrambach Andreas 
Published: May 2005

AbstractThe previous conditions for the physical characterization of turnip crinkle virus (TCV) by quantitative agarose gel electrophoresis [1, 2] were limiting the method to the microgram load level and were therefore insufficiently sensitive to satisfy the need in many areas of virology for detection of viruses containing single‐stranded RNA at the nanogram level. The present report remedies that defect by presenting a technique compatible with the nanogram load level of such viruses. The technique is based on a reduction of gel thickness and on the use of silver staining.
Peptide mapping of polypeptides separated by two‐dimensional electrophoresis: Protease digestion directly on the two‐dimensional gel followed by electrophoresis in reverse direction
Authors: Zivy Michel , Granier Fabienne 
Published: May 2005

AbstractA method is described which allows to reveal simultaneously the proteolytic patterns of numerous polypeptides separated by two‐dimensional electrophoresis. After two‐dimensional electrophoresis, the gels were dipped successively in buffers for preequilibration, protease digestion, and reequilibration. They were then returned to the electrophoresis tank, and electrophoresis was continued for a short time. After silver staining, digestion products appeared, lined up behind the original polypeptide spots. The method allows proteolytic patterns of numerous polypeptides to be viasualized simply and quickly. Among proteins of wheat leaves, 31 groups of related polypeptides were found according to the similarity of their proteolytic patterns.
A simple method for preparation of molecular weight marker proteins for sodium dodecyl sulfate‐polyacrylamide gel electrophoresis by photopolymerizations of hemoglobin subunits
Authors: Sato Hiroshi , Aono Sachiko , Semba Reiji , Kashiwamata Shigeo 
Published: May 2005

AbstractPhotoirradiation of globin, obtained from human hemoglobin, in the presence of oxygen and protoporphyrin produced a series of polymers of globin subunits ranging from dimer to dodecamer. These polymers are useful as molecular weight markers because they cover a relatively wide range of molecular weights (Mr 15 500–186 000) with a constant and narrow interval of Mr 15 500.
Specific antigen detection by immunoelectroendosmosis
Authors: Lewis W. H. P. , Sun K. K. Y. 
Published: May 2005

AbstractA novel approach to the immunological stage of immunoblotting is described. The method employs the phenomenon of electroendosmosis to bring the antibodies sequentially into contact with the antigen. Using a three‐stage peroxidase antiperoxidase system for human group specific protein (Gc) phenotyping, it has been shown that the method has comparable sensitivity, is more rapid and is more economical of antibodies than a similar multi‐stage immerse‐and‐wash system.
Two‐dimensional electrophoresis of human lymphocyte proteins: Two‐dimensional polymorphisms and paternity testing
Authors: Waldinger Dorothea , Cleve Hartwig 
Published: May 2005

AbstractGenetic polymorphisms of seven human lymphocyte proteins, analyzed by two‐dimensional electrophoresis, were evaluated in respect to their suitability for paternity testing. Current data of an enlarged family and population study for five proteins (p23, p30, p40, p60, p66), already described for a smaller population sample of Southern Germany, are presented together with evidence for a new polymorphic protein (p42), recently observed in our survey. These six proteins occurred in isoelectric focusing as two different variants, acidic (a) and basic (b). The genetic basis of the protein variations was ascertained (i) by the presence of homozygous and heterozygous phenotypes, (ii) by the Mendelian mode of transmission of the variants as allelic gene products within 17 families and (iii) by the demonstration of a gene‐dosage dependence comparing the spot intensities in homozygous and heterozygous phenotypes. For quantitative data, laser densitometric scanning of the protein spots followed by computer‐assisted quantitative evaluation of the spot intensities was performed. The allele frequencies of the polymorphic proteins were calculated from the phenotype distributions within a sample of 56 unrelated individuals from Southern Germany. Gene frequencies of the common alleles ranged between 0.991 and 0.518. To discuss the suitability of the two‐dimensional polymorphisms for paternity testing the theoretical exclusion probabilities were assessed for seven polymorphic proteins observed in our population sample, the six polymorphisms with two alleles described here and a further polymorphism (p75) with six alleles. For five proteins (p23, p40, p42, p66 and p75) we found sufficiently high values for the theoretical exclusion probabilities, ranging from 10% to 34%. For the seven polymorphic proteins, which could be analyzed on a single two‐dimensional gel, we calculated a combined theoretical exclusion probability of 67%.
Forensic aspects of haptoglobin: Electrophoretic patterns of haptoglobin allotype products and an evaluation of typing procedure
Authors: Teige Brita , Olaisen Bjørnar , Pedersen Lilian , Teisberg Per 
Published: May 2005

AbstractA procedure used for haptoglobin (Hp) typing in paternity cases has been evaluated. All serum samples have been subtyped with a one‐dimensional isoelectric focusing/immunoblotting method, and samples with rare or questionable patterns have been further examined by two‐dimensional electrophoresis with isoelectric focusing in the first dimension followed by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis in the second dimension. The electrophoretic Hp‐patterns of common and rare α‐ and β‐chain variants are shown, including allotype patterns of two new β‐chain variants and three new α‐chain variants. Retyping of nearly 2000 individuals at intervals between 1 to 12 months revealed a typing error frequency of about 0.3 %, which is considered acceptable, provided new blood samples are required in every case of paternity exclusion. Comparison of typing results obtained with the present procedure and with routine starch gel electrophoresis in more than 5000 serum samples gave conflicting results in 6 samples. The sensivity of the described one‐dimensional subtyping method was slightly better than that of starch gel electrophoresis. In 4110 unrelated individuals, involved in cases of disputed paternity the Hp 2SS 0.038, Hp 2FF 0.004, and Hp 3 (Johnson) 0.0005. These allele frequencies give a theoretical paternity exclusion rate of 32.5%, which is in accordance with the observed rate in 2200 paternity cases with more than 600 non‐fathers. It is concluded that the present procedure represents a definite improvement for Hp subtyping in practical paternity diagnostics. Preliminary results with retyping of weak Hp patterns using a staining technique involving the biotin/avidin complex indicate that the sensitivity of the one‐dimensional subtyping method may be substantially increased. Dried blood stains, containing approximately 1–3 μL blood, have been typed with good results employing the latter staining technique. The applicability of the one‐dimensional subtyping method in forensic casework is presently being investigated.
The group specific component/vitamin D binding protein (GC/DBP) system in the analysis of disputed paternities
Authors: Constans Jacques , Cleve Hartwig 
Published: May 2005

AbstractThe group‐specific component (GC) was discovered in 1959, and in the same year a vitamin D binding protein (DBP) in human plasma was found; however, their identity was established as late as 1975. In the GC/DBP system three common alleles, GC*1F, GC*1S, and GC*2, determine six GC phenotypes: 1F, 1S, 2, 1F‐1S, 2‐1F and 2‐1S, these common alleles having been found in all human populations studied. In addition, more than 120 GC variants have been discovered, with varying frequencies in different populations. The distribution of the common GC phenotypes and the presence of rare GC variant phenotypes render the GC/DBP system useful for the analysis of disputed paternities.
Identification of group specific component/vitamin D‐binding protein (GC/DBP) mutants by isoelectric focusing in immobilized pH gradients
Authors: Constans Jacques , Cleve Hartwig 
Published: May 2005

AbstractThe six common genetic types of the group specific component/vitamin D‐binding protein (GC/DBP) system are usually classified by isoelectric focusing in carrier ampholytes, followed by visualization of the GC proteins by immunoprinting with monospecific antiserum. In addition, more than 120 mutant GC types have been discovered. For their identification additional methods were necessary, including poly acrylamide gel electrophoresis, isoelectric focusing in the presence of 3 M urea as well as isoelectric focusing in immobilized pH gradients. The application of the last method is described in detail and several examples of GC/DBP mutants identified thereby are presented.
Acid phosphatase typing for breeding nematode‐resistant tomatoes by isoelectric focusing with an ultranarrow immobilized pH gradient
Authors: Günther Siegfried , Postel Wilhelm , Wiering Henk , Görg Angelika 
Published: May 2005

AbstractThe genetic variants of tomato acid phosphatase (Aps‐1) systems have been analyzed by isoelectric focusing in immobilized pH gradients (IPG). By using an ultranarrow pH 4.25–4.55 IPG gel, the two genotypes Aps‐11 and Aps‐1+, differentiating tomato variants into nematode‐resistant or nematode‐susceptible plants, are separated into two sharp zones over a distance of 2.5 cm with isoelectric points of 4.37 and 4.43, respectively. Under these conditions, silver staining of the Aps‐1 variants proved to be superior to enzyme staining. By applying more than 50 samples on one IPG gel, this method proved to be a powerful tool for reliable tomato nematode resistance screening.
Electric field gradients and band sharpening in DNA gel electrophoresis
Authors: Slater Gary W. , Noolandi Jaan 
Published: May 2005

AbstractA mathematical study of the effect of non‐uniform electric fields on the width of DNA electrophoretic bands is presented. Using a simple model, we show that field gradients sharpen these bands during an experiment if the corresponding gradient of electrophoretic velocity is large enough. This is in agreement with experimental results indicating that narrower bands form when pulsed field electrophoresis is carried out in the presence of field gradients. Moreover, it is shown that there is in fact an optimal experimental duration that maximizes separation. Finally, gradients are also predicted to reduce the relative mobilities of the DNA fragments, which is a serious drawback of this technique.
Density gradient electrophoresis of cells in a reversible gel
Authors: Plank Lindsay D. , Kunze M. Elaine , Gaines Richard A. , Todd Paul 
Published: May 2005

AbstractDensity gradient electrophoresis permits the separation of cell types according to surface charge density with high resolution. Any source of flow compromises the resolving power of density gradient electrophoresis. Although procedures have been devised to successfully counteract electroosmotic and convective flows, the final collection of separands requires that they be pumped out of the electrophoresis column. Experiments were therefore designed to test the hypothesis that this flow could also be eliminated by trapping the separated bands in a gel, from which they could be collected by slicing the gel cylinder. Glutaraldehyde‐fixed rat and rabbit erythrocytes were used as test particles in a phosphate‐buffered isotonic Ficoll‐sucrose density gradient in a 2.2 cm diameter, thermostated vertical glass column that could be opened at both ends. Two types of agarose were used as gel polymers: Electrophoresis grade agarose (J. T. Baker Chemical Co.) at final concentrations of 0.1 to 0.25 % and SeaPrep ultralow gelling agarose (Marine Colloids Div., FMC Corp.) at a final concentration of 1.0 %. Electrophoretic separability of the test particles and fluid stability were tested independently at 55 °C and 32 °C at which the two agaroses were, respectively, liquid. The experiments demonstrated that the higher temperatures required and the presence of agarose compromised neither the stability of the density gradient nor the migration properties of the cells, and cells can be separated in a sol at a temperature that is compatible with cell viability.
Blotting from PhastGel media after horizontal sodium dodecyl sulfate‐polyacrylamide gel electrophoresis
Authors: Jägersten Christina , Edström Annica , Olsson Birgitta , Jacobson Gunilla 
Published: May 2005

AbstractAn improved procedure, “thermoblotting”, is described for transferring proteins by diffusion from PhastGel™ Gradient media to an immobilizing matrix after horizontal sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. After electrophoresis the gels were left on the separation bed of PhastSystem™, the blotting matrix was applied and a transfer temperature was selected between 5–70 °C. An experimental series at fixed diffusion times showed that the transfer yield was significantly increased with temperature. The evaluation was done visually after staining of the blots with colloidal gold. An evaluation study comparing nitrocellulose, nylon, and polyvinyl‐idenedifluoride of different pore sizes is also reported. Finally, the transfer efficiencies for 125I‐labelled bovine serum albumin and soybean trypsin inhibitor were estimated using four different blotting procedures: two diffusion blotting techniques and two electrophoretic blotting techniques (tank vs. semi‐dry).
An evaluation of two‐dimensional electrophoresis for detection of human serum proteins in acute myocardial infarction
Authors: Marshall Thomas , Williams John , Williams Katherine M. 
Published: May 2005

AbstractHigh resolution two‐dimensional electrophoresis indicates that serum proteins previously only detected in patients with acute myocardial infarction (AMI) (Gomo et al., Electrophoresis 1983, 4, 298–302) are also present in the serum protein patterns of other patients (AMI negative or demonstrating acute phase response) and are faintly detected even in controls. Thus, these proteins are not specific to AMI and are probably acute phase reactants. However, they do demonstrate a characteristic time course response in sequential samples from AMI patients.
Two‐dimensional electrophoresis with immobilized pH gradients of leaf proteins from barley (Hordeum vulgare): Method, reproducibility and genetic aspects
Authors: Görg Angelika , Postel Wilhelm , Domscheit Albert , Günther Siegfried 
Published: May 2005

AbstractLeaf proteins from 14 barley cultivars (Hordeum vulgare) were analyzed by two‐dimensional electrophoresis with immobilized pH gradients (IPG 4–7 and IPG 6–10) in the first dimension. Highly reproducible two‐dimensional patterns were obtained, owing to constant spot positions along the isoelectric focusing axis. A number of variety‐specific protein spots were detected, allowing us to discriminate barley cultivars not only into main groups but into individual cultivars.
The use of two‐dimensional gel electrophoresis in the analysis of organ‐specific maize proteins
Authors: Goday Adela , Torrent Margarita , Ludevid M. Dolors , Puigdomènech Pere 
Published: May 2005

AbstractTwo‐dimensional gel electrophoresis with non‐equilibrium pH gradient electrophoresis in the first dimension and sodium dodecyl sulfate‐polyacrylamide gels in the second dimension has been used for the analysis of organ‐specific proteins in maize. The method has been used to study the whole protein pattern of developing organs and adult leaves as well as protein patterns of in vitro translation. Examples of two‐dimensional immunoblotting and in vitro translation of endosperm‐specific proteins are also shown. Two‐dimensional gel electrophoresis appears as an essential analytical step in the identification of organ‐specific proteins and for the detection of the protein products related to organ‐specific genes identified by other means.
Genetic aspects of variation of protein amounts in maize and pea
Authors: de Vienne Dominique , Leonardi Agnès , Damerval Catherine 
Published: May 2005

AbstractUsing high‐resolution two‐dimensional polyacrylamide gel electrophoresis we studied the polymorphism of protein amounts in some genotypes of maize and pea. This type of variability seems to be rather common and insensitive to environmental conditions, as attested by the comparison of the patterns of two maize lines harvested in two different years. A large‐scale experiment involving 5 lines, 7 of their hybrids, and 6 organs (or physiological stages) of maize allowed us to examine numerous polypeptides regrading their genetic variability, their amount differences between organs and the inheritance of their abundance. Genetic and organ variations are not independent: polypeptides whose amount varies from one organ to another are, for the most part, genetically variable (59 %), while the stable polypeptides are not often genetically variable (18 %). We found a striking organ specificity for (i) the extent of quantitative variability (from 2.3–15.4 % of the polypetides), (ii) the occurrence and the type of variation for a given polypeptide (an intensity difference seen in an organ can disappear or even be reversed in another one), (iii) the kind of inheritance (additive/non‐additive): combining the 6 organs and the 7 hybrids we found 101 cases of non‐additivity (4 % of the total) which conecern as many as 72 different spots, that is to say that in most cases a polypeptide displaying nonadditivity in an organ seems to display additivity in the other ones. Moreover, for most of the polypeptides with nonadditive inheritance the hybrid spot presents an intensity similar to that of the most intense parental spot. Thus for protein experssion a hybrid cannot be predicted from its parents. The organ‐specificity was also evidenced by analyzing pleiotropic effect of gene substitutions in various organs of garden pea. Comparisons of isogenic lines differing for the r‐locus revealed that according to the organ, the gene substitution caused appearances, disappearances or quantitative changes for 0 % to more than 10 % of the proteins revealed. All these features are discussed in connection with the possible role of variation of protein amounts in phenotypic variability.