bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization and visualization of the low density lipoprotein (LDL) receptor by ligand blotting using anti‐LDL‐enzyme linked immunosorbent assay
Authors: Dresel Hans A. , Schettler Gotthard 
Published: May 2005

AbstractThis paper describes the visualization of the low density lipoprotein (LDL) receptor by ligand blotting technique using an anti‐LDL‐enzyme linked immunosorbent assay (ELISA). Solubilized membrance proteins from bovine adrenal cortex are subjected to sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis, transferred to nitrocellulose paper and incubated with LDL. The combination of peroxidase‐con‐jugated second antibody and 4‐chloro‐naphthol/H2O2 allows rapid development of colored bands where LDL was bound to the receptors. Saturation of the LDL binding to the LDL receptor can be demonstrated. This highly sensitive and rapid ELISA avoids the need for continuous supplies of iodinated antibodies.
Non‐linear pH courses with immobilized pH gradients
Authors: Gianazza Elisabetta , Giacon Patrizia , Sahlin Bengt , Righetti Pier Giorgio 
Published: May 2005

AbstractExamples of increased resolution obtained with non‐linear pH gradients are presented. Discussed in detail is the “ideal” pH course in the range 4–10, based on the statistical distribution of isoelectric points of protein from data in the literature.
Cell electrophoresis: Automatic measurements by laser light scattering with Lazypher
Authors: Steiner Rudolf , Ottmann Oliver , Kaufmann Raimund , Ann Light P. , Hoffmann Walter 
Published: May 2005

AbstractAutomated cell electrophoretic measurements with Lazypher are based on the laser Doppler technique. Experiments are performed with high precision (of 1.2% S. D.) and in short time intervals of 1–3 min. Application to electrophoretic mobility tests with tanned sheep erythrocytes incubated in supernatants from sensitized murine thymus and spleen cells which had been prepared from cultures containing 5 μg purified protein derivative (PPD)/ml caused a reduction of the electrophoretic mobility (EM) of 5% and 13%, respectively. Control spleen cell supernatants caused a 4% decrease in mobility and no effect was observed with supernatants from control thymus cells. Peripheral blood lymphocytes from healthy adults (n = 56) and children with atopic diseases and congenital heart disease (n = 12) are separated into a high mobility peak (μ = 1.24 μmcm/Vs for adults, μ = 1.19 μmcm/Vs for the child group) corresponding to T‐cells and a low mobility peak, μ = 0.93 μcm/Vs, representing B‐cells. The mobility difference between the adult and child T‐cells is highly significant (99%). Electrophoretic measurements of myeloid cells from healthy donors and patients with chronic myeloid leukemia (CML) and acute myeloid leukemia (AML) show that progressing differentiation within this cell line is accompanied by a decreasing mobility. Blast cells from a patient with acute lymphoblastic leukemia (c‐ALL) had an EM, μ = 1.09 μmcm/Vs, distinct from that of myeloblasts in AML (μ = 1.16 and 1.25 μmcm/Vs) and CML (μ = 1.25 and 1.23 μmcm/Vs) and from normal T‐and B‐lymphocytes.
Some more formulations for immobilized pH gradients
Authors: Gianazza Elisabetta , Astrua‐Testori Silvia , Righetti Pier Giorgio 
Published: May 2005

AbstractFormulations are presented for 4 to 6 pH unit wide immobilized pH gradients requireing the use of only six available Immobiline buffers, without strong acidic and basic titrants. Recipes are also given for the pH 3–10 range.
Computer automated lanes detection and profiles evaluation of one‐dimensional gel electrophoretic autoradiograms
Authors: Pun Thierry , Trus Benes , Grossman Nili , Leive Loretta , Eden Murray 
Published: May 2005

AbstractA new fast, precise and reproducible method for lanes detection and profiles evaluation of one‐dimensional gel electrophoretic autoradiograms is presented. It is fully automated, and its principles are based on image and signal processing techniques. The operator usually has only to specify the number of lanes that are present in the preparation, but still may retain overall control. In the first phase, the program automatically locates the lateral position of the lanes, and approximates their left and right boundaries by polynomials (which allow curved as well as straight lanes). The operator can then intervene, if necessary, to adjust these positions. In the second phase, the background variations are corrected; two types of density profiles are computed, representing the average density as well as the total amount of material in the lanes. This calculation is carried out with correction for tilt or curvature for each lane. Finally, prinouts are produced that contain graphs of the profiles together with quantitative measurements. Use of this method is illustrated by analysis of electrophoretic gels containing proteins and lipopolysaccharides of varying mobilities. Experiments made on these gels as well as others yield accurate quantitation of the relative amount of each component.
A simple method for drying of sodium dodecyl sulfate‐polyacrylamide disc gels
Authors: Aono Sachiko , Sato Hiroshi , Semba Reiji , Kashiwamata Shigeo 
Published: May 2005

AbstractA simple method for the drying, on filter paper, of sodium dodecyl sulfate polyacrylamide disc gels is described. This method has been successfully applied to disc gels with polyacrylamide concentrations of 5, 10, and 15%, and also with a linear gradient of 5–15%. The gels dehydrated without any longitudinal shrinkage are suitable for longtime storage and may be used for molecular weight estimation of proteins.
Effect of conductivity and concentration on the sample stream in the transverse axis of a continuous flow electrophoresis chamber
Authors: Miller Teresa Y. , Williams George O. , Snyder Robert S. 
Published: May 2005

AbstractThe resolution of continuous flow electrophoresis systems is generally measured by the spread of the sample bands in the direction of the electrophoretic migration. This spreading assumes an electric field that extends uniformly between the two electrodes. This paper evaluates the cross‐section of the sample bands in the plane perpendicular to the flow and shows that the spread in the direction perpendicular to the migration increased significantly with the applied electic field. Concentrated samples of monodisperse latex particles and vinyltoluene T‐butylstyrene particles in sample buffers of different electrical conductivities were used to map the shape of the sample bands relative to the zero electric field case. As the electric field was applied, the sample band spread from an initial diameter of only one‐third the chamber thickness until it approached the chamber walls where electroosmosis significantly reduced the resolution of separation. It can be shown, however, that it is possible to minimize these distortions by careful sample preparation and experiment design.
Differences in two‐dimensional patterns of cellular proteins from murine T‐ and B‐lymphocytes after mitogenic stimulation
Authors: Braun Andreas , Waldinger Dorothea , Cleve Hartwig 
Published: May 2005

AbstractSplenocytes from inbred mice strains were stimulated with concanavalin A (Con A) or lipopolysaccharide (LPS) to form T‐ or B‐blasts. Following three days of mitogenic stimulation, immunofluorescence of the splenocytes with a rabbit antimouse‐immunoglobulin antiserum conjugated with fluorescein isothiocyanate revealed an extensive enrichment of T‐blasts by Con A and of B‐blasts by LPS. The cell preparations, however, represented enriched but not pure populations of T‐ or B‐blasts. Two‐dimensional electrophoretic resolution of cellular proteins from Con A or LPS‐stimulated murine splenocytes and subsequent silver staining of the polyacrylamide gels showed approximately 450 spots (polypeptides). Comparative analysis of the resulting patterns revealed 13 differences specific to the T‐cells and 8 specific to the B‐cells. In order to ascertain the reproducibility of the differences of two‐dimensional protein patterns from Con A and LPS stimulated splenocytes, the experiment was performed several times using two different inbred mice strains (C57B1/6 and BALB/c). Interstrain specific differences were also found at a molecular mass of about 68 kDa and at a pI range between 6.1 and 5.85.
Genetic polymorphism of human serum transferrin in the swiss population: Evidence for three “new” transferrin‐variants
Authors: Scherz Rudolf , Reber Beatrice , Pflugshaupt Rolf , Bütler René 
Published: May 2005

AbstractGenetic polymorphism of transferrin (Tf) was studied in 473 healthy Swiss blood donors and 236 nonrelated individuals engaged in cases of disputed paternity. Gene frequencies were: TfC1 = 0.7727, TfC2 = 0.1634, TfC3 = 0.0573. Several rare alleles were observed, the frequency of the total of which was 0.0066. Three obviously new Tf alleles were detected. The phenotype distribution was in agreement with the Hardy Weinberg equilibrium. Among 113 mother/child pairs no phenotypes contradictory to the mendelian law were observed. The transferrin system represents a useful parameter in routine paternity testing. Its average chance of exclusion (without rare alleles) is 18.19%.
Quantitative reproducibility of measurements from Coomassie Blue‐stained two‐dimensional gels: Analysis of mouse liver protein patterns and a comparison of BALB/c and C57 strains
Authors: Leigh Anderson N. , Nance Sharron L. , Tollaksen Sandra L. , Giere Frederic A. , Anderson Norman G. 
Published: May 2005

AbstractUsing the ISO‐DALT system for two‐dimensional (2‐D) electrophoresis and the TYCHO system for computer analysis of the resulting protein maps, we obtained high quality quantitative protein abundance data from Coomassie Brilliant Bluestained gels of mouse liver samples. High resolution gels allow more than 100 proteins to be measured with coefficients of variation less than 15 %. A comparison of results from two mouse strains (C57BL/6 and BALB/c) and the cross between them (BCF1) shows that a large number of qutive polymorphisms can be detected, and that, as expected, the amount of protein produced in the heterozygote is intermediate between the parental values. The system described is shown to be capable of reliably detecting decreases in protein abundance such as those expected to result from radiation‐induced deletion of one copy of a gene. The implications of these results for the study of gene regulation are discussed in relation to applications in genetics, toxicology, and differentiation.
Electrophoretic analysis of the glycan microheterogeneity of orosomucoid in cancer and inflammation
Authors: Hansen John‐Erik S. , Jensen SøRen P. , Nørgaard‐Pedersen Bent , Bøg‐Hansen Thorkild C. 
Published: May 2005

AbstractUsing crossed affinoimmunoelectrophoresis with free concanavalin A the glycandependent microheterogeneity of the immunoregulatory glycoprotein orosomucoid was examined in serum from 30 healthy individuals, 28 patients with benign chronic inflammation and 56 patients with cancer of various types and stages and before and after surgery. We found significant differences in glycan structure between the different patient groups. Thus the patients with the acute phase reaction of surgery differed from patients with chronic inflammation, and patients with testicular cancer differed from patients with colorectal cancer in having a greater proportion of biantennary glycan orosomucoid. Surgery was found to result in a temporary change in the relative proportions of the three microheterogeneity types of orosomucoid, lasting about one week.
New assay procedure for glutathione S‐transferase using analytical capillary isotachophoresis: A procedure for the simultaneous analysis of reduced and oxidized glutathione together with glutathione conjugates of electrophilic compounds
Authors: Holloway Christopher J. , Battersby Rüdiger V. 
Published: May 2005

AbstractA procedure for the simultaneous analysis of reduced and oxidized glutathione together with glutathione conjugates of electrophilic organic compounds by capillary isotachophoresis is described. This method can be applied to quality control of such synthetic conjugates, which are commonly employed as affinity ligands in the chromatographic purification of glutathione S‐transferases (EC 2.5.1.18). Using this new assay procedure, it has been found that the glutathione conjugate of 1‐chloro‐2,4‐dinitrobenzene is unstable in neutral to alkaline solution, degrading to glutathione and 2,4‐dinitrophenol. Anomalies in the electrophoretic mobilities of a homologous series of alkylglutathione derivatives were observed. A possible explanation is provided on the basis of the formation of micelle‐like aggregates, this tendency increasing with increasing length of the hyrocarbon chain. A useful and widely applicable aspect of this analytical procedure is as a method of monitoring glutathione S‐transferase. The major advantages of this new assay technique can be summarized as follows: (1) the assay does not depend on optical properties of the acceptor, (2) the reaction can be monitored in the presence of unrefined biological material;aliquots can be injected without pretreatment and (3) the spontaneous oxidation of glutathione in the assay mixture can be monitored simultaneously.
Influence of the state of denaturation on the migration of adenovirus type 2 structural proteins in sodium dodecyl sulfate polyacrylamide gels
Authors: Caillet‐Boudin Marie L. , Lemay Pierre 
Published: May 2005

AbstractThe changes in the migration of adenovirus type 2 structural proteins in sodium dodecyl sulfate polyacrylamide gels following various denaturation conditions or periods have been studied. Several factors such as proteolytic cleavages, disulfide bridges, carbamylation or residual charge effects which could lead to altered migration were screened. Resistant high order structures sensitive to the action of urea might cause these variations. A likely relationship between the difference of apparent molecular weight, frictional coefficient and hydrophobicity has been established.
The use of free flow electrophoresis in the purification of recombinant human tissue plasminogen activator expressed in yeast
Authors: Barth Françoise , Gruetter Markus G. , Kessler Reinhard , Manz Hans‐Jörg 
Published: May 2005

AbstractFree flow electrophoresis was used for the purification of human tissue plasminogen activator (t‐PA) produced by recombinant yeast cells. This method was employed for three principal reasons: (a) yeast t‐PA purified by immunoaffinity chromatography was not pure enough to carry out protein chemical characterization, (b) the amount of yeast t‐PA available for purification was extremely limited, (c) t‐PA has a strong tendency to adsorb to support material; a support‐free separation method therefore seemed to be a reasonable way to further purify the enzyme. Free flow electrophoresis was first performed using solutions containing the soluble proteins of yeast cell extract, and secondly, using t‐PA fractions prepared by immunoaffinity chromatography, Yeast t‐PA purified by immunoaffinity chromatography and then by free flow electrophoresis is more than 80% pure and suitable for characterization experiments such as glycosylation studies or N‐terminal sequence determination.
Analysis of polymeric proteins by two‐dimensional polyacrylamide gel electrophoresis on one slab in the presence and absence of sodium dodecyl sulfate
Authors: Bloemendal Hans , Jansen Kees 
Published: May 2005

AbstractThe method presented was developed specifically to be used for the electrophoretic analysis of polymeric proteins in order to establish the number of aggregates and that of their subunits. Electrophoresis is carried out on a single slab; without sodium dodecyl sulfate (SDS) in the first dimension and with SDS in the second dimension. The method is possible since for one‐dimensional SDS gel electrophoresis, SDS is only required in the cathode vessel.
2‐Pyridinecarboxylate as counter ion in capillary isotachophoresis of low‐mobility bases
Authors: Jokl Vladimír , Pospíchalová Jaroslava , Polášek Miroslav 
Published: May 2005

AbstractAn electrolyte system involving 2‐pyridinecarboxylate (Pic) buffer as the leading and formic acid as the terminating electrolyte was suggested for cationic capillary isotachophoresis of bases with low mobilities. Parameters of the system were studied for concentrations of the leading K+ ion of 1–30 mmol/L and for the Hpic/Kpic ratios of 3:1, 1:1 and 1:3 in the buffer. The effective mobility of the terminating H+ ion ranged between 3 and 30 X 10−9 m2V−1s−1 depending on the composition of the electrolyte system. The applicability of the system (at optimum concentration of K+ 5 mmol/L and the HPic/Kpic ratio 1:1) to the analysis of bases was characterized by a zone existence diagram and demonstrated by the separation of a model mixture containing tetramethylammonium, tetraethylammonium, tris(hydroxymethyl)aminomethane, ephedrine, codeine, nortriptyline, 1,10‐phenanthroline, aminophenazone and 4‐aminoantipyrine.
Ultrathin‐layer sodium dodecyl sulfate‐polyacrylamide gradient gel electrophoresis and silver staining of urinary proteins
Authors: Schiwara Hans‐Walter , Hebell Thomas , Kirchherr Hartmut , Postel Wilhelm , Weser Johannes , Görg Angelika 
Published: May 2005

AbstractUltrathin‐layer sodium dodecyl sulfate‐polyacrylamide gradient gel electrophoresis in combination with silver staining is a highly sensitive and rapid method for molecular size analysis of urinary proteins. First results are achieved after 4 h (electrophoretic run, 100 min, followed by Coomassie Brilliant Blue staining, 2 h). After additional silver staining, including a recycling step, the results are ready for evaluation within 7 h. Urine samples do not need to be concentrated. Within a single gel, 25–27 samples can be run side by side under identical conditions and thus compared without ambiguity. By coelectrophoresis of pure proteins and immunological analysis 13 urinary proteins could be identified and for 7 of them the detection limits were determined. Glomerular protein patterns in samples of urine with a protein concentration in the normal range are compared with the concentration of albumin and transferrin measured by a sensitive enzyme immunoassay. Typical renal and extrarenal protein patterns are shown.
Free‐flow electrophoresis in solutions of borax and mannitol
Authors: Shukun Sergei A. , Gavryushkin Alexander V. , Brezgunov Vyacheslav N. , Zav'yalov Vladimir P. 
Published: May 2005

AbstractFree‐flow electrophoresis of proteins in solutions of borax and mannitol was investigated. The borax solution represents an equimolar boric acid‐NaOH buffer with a pH of 9.2. On addition of mannitol the pH of the buffer decreases, with increasing polyol concentration, accompanied by a slight decrease in conductivity. Using a 2.5 mM borax solution, buffers with pH from 9.2 to 4.5 can be obtained. Under the influence of the electric field the pH and conductivities of these buffers change in the near‐membrane regions of the separation chamber. Despite the changes the buffers are suitable for electrophoresis of proteins in a free‐flow apparatus. The separation of bovine hemoglobin as well as a mixture of bovine hemoglobin, human serum albumin and cytochrome C in borax‐mannitol solutions is demonstrated at 80–100 mA constant current and a residence time of only 2.0–2.6 min.
Identity of Treponema pallidum subsp. pallidum polypeptides: Correlation of sodium dodecyl sulfate‐polyacrylamide gel electrophoresis results from different laboratories
Authors: Norris Steven J. , Alderete John F. , Axelsen Nils H. , Bailey Mark J. , Baker‐Zander Sharon A. , Baseman Joel B. , Bassford Philip J. , Baughn Robert E. , Cockayne Alan , Hanff Philip A. , Hindersson Peter , Larsen Sandra A. , Lovett Michael A. , Lukehart Sheila A. , Miller James N. , Moskophidis Matthäus A. , Müller Ferdinand , Norgard Michael V. , Penn Charles W. , Stamm Lola V. , van Embden Jan D. , Wicher Konrad 
Published: May 2005

AbstractAs the first step in a cooperative effort to standardize the identification of the polypeptides of Treponema pallidum subsp. pallidum, the sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) results obtained in 16 laboratories were compared. Although it was possible to correlate the positions of 16 of the major polypeptide bands, the cross‐identification of many of the polypeptides was ambiguous, particularly in the low molecular weight range. Two‐dimensional electrophoresis provided an improved means of separating and characterizing T. pallidum polypeptides as isolated molecular species. An approach to the unambiguous identification of treponemal polypeptides was outlined which will utilize two‐dimensional electrophoresis in combination with specific properties attributable to individual proteins, including reactivity with monoclonal antibodies or monospecific antisera, biochemical and structural properties, and sequence information. To demonstrate the feasibility of this approach, two‐dimensional electrophoresis in conjunction with immunoperoxidase staining was used to specifically identify three cloned T. pallidum proteins.
Application of two‐dimensional electrophoresis in comparing the effects of progestins and tamoxifen on proteins of the abnormal uterus
Authors: Guest Julian F. , Elder Murdoch G. , White John O. 
Published: May 2005

AbstractIn the androgenized rat, uterine histological abnormalities associated with unopposed oestrogenic stimulation can be partially reversed following progestin therapy. The effects of progestins and tamoxifen on the uterine protein composition of androgenized animals have therefore been investigated in an attempt to identify hormonally sensitive proteins that reflect changes in tissue morphology. Subcellular fractions prepared from uteri of androgenized animals were analysed by one‐ and two‐dimensional gel electrophoresis. In all fractions, protein changes were observed and the identity of some of these proteins has been tentatively assigned following peptide mapping. Evidence is presented of an apparent correlation between potency to antagonize the effects of unopposed oestrogenic stimulation and an increase in the proportion of cytosolic glucose‐6‐phosphate dehydrogenase and the B form of creatine kinase.
Group‐specific component subtyping: A high resolution method employing short run cathodic drift
Authors: Kuo Sze‐Ern 
Published: May 2005

AbstractA high resolution polyacrylamide gel isoelectric focusing method is described for group‐specific component (Gc) subtyping. With this method all the common Gc subtypes can be identified and the Gc variant 1A1 bands are clearly resolved from the Gc 1F and 1S bands. The run time of 90 min represents a substantial saving in time when compared to many other methods reported in literature.
Anti‐mitochondrial protein antibodies in a serum from a patient with systemic lupus erythematosus: Specificity and comparison with other anti‐mitochondrial antibodies
Authors: Celis Julio E. , Madsen Peder , Andersen Irene , Andersen Paul , Vayssiere Jean‐Luc , Croizat Bernard , Thiessen Hanne , Nielsen Søren 
Published: May 2005

AbstractAnti‐mitochondrial antibodies present in a serum from a systemic lupus erythematosus patient have been characterized by indirect immunofluorescence of cultured transformed human amnion cells, and by two‐dimensional gel electrophoresis and immunoblotting. The antibodies which stained mostly mitochondria in transformed human amnion cells reacted with four mouse brain cortex mitochondrial proteins. These proteins, which have been termed a, b, c and d, exhibited apparent molecular weights of 66 000, 49 000, 49 000 and 44 000, respectively. Proteins a and c exhibited charge variants. Surprisingly, most of the proteins recognized by this serum also reacted with sera from a patient with liver cirrhosis and idiopathic pulmonary fibrosis.