bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fabric‐reinforced ultrathin polyacrylamide gels for sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and isoelectric focusing
Authors: Peisker Klaus 
Published: May 2005

AbstractThe suitability of four different fabric materials for the preparation of ultrathin reinforced polyacrylamide gels was investigated. With all fabric‐reinforced gels, a good separation of proteins by isoelectric focusing and sodium dodecyl sulfate‐electrophoresis could be achieved. Semi‐dry electrophoretic blotting of proteins was possible with all types of fabric‐reinforced gels. Two polyester fabrics (a net and a fleece) were decidedly superior in handling and dimensional stability on drying to a nylon fabric and another polyester fleece material. Only gels prepared with the former materials withstood further treatment, such as fixation, staining, destaining, and drying. One of the polyester fleece fabrics had poor handling properties and the nylon fabric was unsuitable for direct staining procedures employing concentrated (20 % w/v) trichloroacetic acid as fixative.
Degradation of individual intracellular proteins analyzed by two‐dimensional gel electrophoresis and computerized video densitometry
Authors: Rodemann H. Peter 
Published: May 2005

AbstractA technique is described for the analysis of degradation rates of individual intracellular proteins, based on pulse‐chase‐labeling of cells using radioactive amino acids [35S]methionine, two‐dimensional polyacrylamide gel electrophoresis, fluorography and scanning of the fluorograms by a computerized video densitomter. As compared to scintillation counting of individual protein spots resolved by two‐dimensional gel electrophoresis, this method allows a rapid and precise determination of the degradation rates of individual intracellular proteins. In the present study, degradation rates of individual intracellular proteins of normal human skin fibroblasts and skin fibroblasts from patients with Duchenne muscular dystrophy were compared. Rates of degradation for proteins PIIa, PIIb and PIIc recently described as cell‐type‐specific proteins were significantly enhanced (p > 0.01) in fibroblast cultures of Duchenne muscular dystrophy origin.
Electrophoresis in an expanded stationary boundary
Authors: Michov Budin M. 
Published: May 2005

AbstractA method of electrophoresis in an expanded stationary boundary is described, in which a pH gradient formed in a single buffer is used. The degrees of ionization and the concentrations of the electrolytes have values at which the polyions stack and resolve simultaneously in a polyacrylamide gel. To illustrate the method, the resolving of serum proteins in a pH gradient formed by Tris – glycinate buffer is considered.
Free flow electrophoresis for the purification of proteins: I. Zone electrophoresis and isotachophoresis
Authors: Hoffstetter‐Kuhn Sabrina , Kuhn Reinhard , Wagner Horst 
Published: May 2005

AbstractThe principles and some applications of free flow zone electrophoresis and isotachophoresis are described. The influence of (i) carrier electrolyte conductivity on the migration velocity and (ii) band shape on zone electrophoresis was investigated. The technique was found convenient for studying the effect of pH on the mobility of proteins to create a mobility curve. The purification of alcohol dehydrogenase from a crude yeast extract revealed the separation power of zone electrophoresis for complex protein mixtures. Without additional steps, a purification factor of 5.4, with a recovery of 97 % alcohol dehydrogenase, was achieved. Free flow isotachophoresis was applied to the purification of immunoglobulins from human serum. Disadvantages of this technique are the time‐consuming development of an optimized separation system and the empirical search for suitable spacers. Also, reaching of the steady state becomes increasingly difficult as the number of sample components increases.
Electrophoresis of gliadins in long acrylamide gels: Method and nomenclature
Authors: Branlard Gérard , Picard Brigitte , Courvoisier Christian 
Published: May 2005

AbstractGliadins, defined as wheat kernel storage proteins soluble in 70% ethanol, possess an electrophoretic diversity permitting us to identify different varieties. Because of genetic proximity, however, it sometimes proves impossible to distinguish between different varieties using standard methods. An electrophoresis method, utilizing a discontinuous buffer system with aluminum lactate and potassium lactate in the gel and sodium lactate in the electrode vessels, is described for two types of gels 18 cm and 32 cm in length. The number of bands in higher than with standard methods, and is significantly increased with long gels, facilitating the distinction between varieties. A nomenclature for the new bands is presented for several widely different cultivars.
Haptoglobin subtyping by isoelectric focusing in miniaturized polyacrylamide gels rehydrated in presence of 2‐mercaptoethanol
Authors: Alonso Antonio , Visedo Guillermo , Sancho Manuel , Fernández‐Piqueras José 
Published: May 2005

AbstractA fast isoelectric focusing method for routine haptoglobin (Hp) subtyping is presented. This method is based on isoelectric focusing, under reducting conditions, of neuraminidase‐treated plasma samples by using dry miniaturized (interelectrode distance: 55 mm) polyacrylamide gel, rehydráted in presence of 2‐mercaptoethanol and a mixture of pharmalyte carrier ampholytes (pH 4–6.5 and pH 6–8) followed by immunoblotting. The presence of 2‐mercaptoethanol in the gel prevented refolding of the Hp α and Hp β chains during focusing, making it possible to obtain a sharp Hp band pattern with a clear separation of the different Hp α allelic products (1S, 1F, 2FS, 2SS and 2FF). A population study carried out with 250 unrelated individuals living in Central Spain is also presented.
Human skeletal muscle myosin light chains analyzed by immobilized pH gradients during ontogenesis: Identification of new phosphorylatable isoforms of light chain 2
Authors: Pernelle Jean‐Jacques , Righetti Pier Giorgio , Wahrmann Juan P. , Herve Bernadette 
Published: May 2005

AbstractPrevious studies using two‐dimensional gel electrophoresis have described adult and fetal isoforms of skeletal muscle myosin light chains (MLC). They have also revealed an embryo‐specific light chain (LC1emb), apparently absent in most adult skeletal muscles. In order to characterize more thoroughly the MLC family, we have analyzed the MLCs from human skeletal muscle at different developmental stages using a two‐dimensional electrophoresis technique with an immobilized pH gradient in the first dimension. The high resolution of this novel technique, resolving components which in isoelectric points are ≤ 0.01 pH, combined with sensitive silver staining, has allowed us to identify four phosphorylatable isoforms of MLC2: two slow‐myosin light chains (MLC2Sa and b), two fast myosin light chains (MLC2:a and b), and their phosphorylated counterparts: MLC2SaP and bP, MLC2FaP and bP. The following major modifications during development were observed: (i) The embryonic LC (LC1 emb) persists up to at least 26 weeks of fetal life. (ii) The polymorphism of LC2 is already evident at 10 weeks of development but only the nonphosphorylated forms of LC2S and LC2F seem to be present. The LC2Fa form is predominant. As early as 26 weeks of fetal life, the 4 phosphorylated forms are detected. In the adult, LC2Fb is a minor component. (iii) LC3F (fast) is already expressed at an early embryonic stage (10 weeks).
Abnormal protein spots revealed by two‐dimensional electrophoresis in mycoplasma‐infected human fibroblasts
Authors: Yang Rei‐Cheng , Tsuji Akihiko , Suzuki Yoshiyuki 
Published: May 2005

AbstractSix new protein spots were detected by two‐dimensional electrophoresis in cultured human fibroblasts infected with mycoplasma. No changes were observed in other protein spots present in normal and pathological cells. After treatment with an antimycoplasma drug for a week, the new spots disappeared and the cells became negative for mycoplasma stain.
Autoradiography using storage phosphor technology
Authors: Johnston Richard F. , Pickett Siobhan C. , Barker David L. 
Published: May 2005

AbstractWe describe the application of photostimulable storage phosphor imaging plates to autoradiography of samples labeled with 32P, 14C and 35S. Imaging plates can detect a 32P‐labeled sample at an exposure level of 1 disintegration/mm2, and 14C and 35S at 25 disintegrations/mm2. Compared to X‐ray film, imaging plates offer increased sensitivity of 15‐ to 250‐fold for 32P and 20‐ to 100‐fold for 14C and 35S. Resolution is in the 0.3 mm range and is therefore suitable for any gel or blot application.
The application of schlieren optics for detection of protein bands and other phenomena in polyacrylamide gel electrophoresis
Authors: Takagi Toshio , Kubota Hidehiro 
Published: May 2005

AbstractA modernized schlieren optics system was applied to follow protein bands visually during polyacrylamide gel electrophoresis. This approach is suitable for quantitative measurement of electrophoresis as well as for detection of protein bands because of the absence of distortion of the electrophoretic pattern during the staining process. The optics are also suitable for the detection of various background phenomena which usually escape observation.
Quantitative autoradiographic imaging using gas‐counter technology
Authors: Bateman J. Edmond 
Published: May 2005

AbstractGas counters based on the multiwire proportional counter (MWPC) and multistep avalanche (MSA) technologies can combine the functions of imaging and quantitation (or assay) of radioactively labelled electrophoretograms and blots. Submillimeter spatial resolutions combined with good quantitation have been achieved for the most common beta‐emitting radio labels. The current state of development of this technology is reviewed and assessments of the performance of these devices relative to the more traditional systems (X‐ray film and the scintillation counter) presented. Particular attention is paid to the potential of these devices to enhance the productivity of the scanning of blots and the performance of assays.
Separation of durum wheat proteins by ultrathin‐layer isoelectric focusing: A new tool for the characterization and quantification of low molecular weight glutenins
Authors: Morel Marie‐Hélène , Autran Jean‐Claude 
Published: May 2005

AbstractAn isoelectric focusing method capable of resolving all groups of storage protein of the wheat seed, including the most basic low molecular weight glutenin (LMWG), was developed. Ultrathin polyacrylamide gels were used after drying and rehydration with 8 m urea, 50 mm DTE and 2.4 % carrier ampholytes (pH 4–9). Densitometric scanning of the isoelectric focusing gels permitted a more accurate and specific quantitation of LMWG components among various cultivars than patterns based on sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. The two main genetic types (i. e. ‘γ‐42’ and ‘γ‐45’) of durum wheats were separated on the basis of the proportion in LMWG in storage proteins, but no significant difference was found within these groups. Advantages of the system as regards reliability, high resolution ability to abolish protein oxidation and preventing reaggregation of LMWG were also discussed.
Quantitative densitometry from the point of view of information theory
Authors: Pardowitz Iancu , Ehrhardt Wolfgang , Neuhoff Volker 
Published: May 2005

AbstractThe process of quantitative densitometry is analyzed with methods developed in information theory. It is shown that the steps involved in densitometry, e. g. gel staining, mechanical, optical and electronic processing, as well as all the steps of data processing, can be viewed as communication channels. The factors affecting both the relevant and irrelevant part of the total information passed through these channels are discussed in the consistent frame provided by information theory. This view leads to a unifying context for analyzing the performance of quantitative densitometers.
GELANAL, a personal computer‐program to compare protein patterns on two‐dimensional polyacrylamide gels
Authors: Klerk Hans , Jespers Ad 
Published: May 2005

AbstractComparing and analyzing a series of two‐dimensional gels by hand is troublesome and subjective. So far a number of systems for automatic analysis have been developed on mainly mainframe computers, using complex algorithms. This paper presents an inexpensive system, based on a simple Pascal program, to compare individual spots on two‐dimensional gels using an IBM or compatible personal computer in a qualitative way. The accuracy of the method is demonstrated by comparing two patterns of the same extract from different runs.
Haptoglobin subtyping with anti‐haptoglobin α chain antibodies
Authors: Shindo Shoko 
Published: May 2005

AbstractSpecific antibodies against the human haptoglobin (HP) α chain were raised and used for immunoblotting after isoelectric focusing to determine Hp α subtypes in a reproducible and simplified manner. By eliminating the staining of the HP β chain, HP α subtypes were visualized more precisely and simply than by previously reported methods. Subtypes of a total of 1211 sera, obtained from two different populations in Japan, were examined by the new method. Four HP α subtypes (HP 1S–1S, 2FS–1S, 2FS–2FS and 2FS–2SS) and five subtypes combined with a new variant (HP 2FS–1V1, 2V1–1S, 2FS–2V1, 2FS–2V2 and 2FS–2V3) were observed. HP 1V1 belonged to HP α1, and HP 2V1, 2V2 and 2V3 belonged to HP α2, and their alleles were designated HP A*1V1, HP A*2V1, HP A*2V2 and HP A*2V3, respectively. The allele frequencies were calculated to be as follows: HP A*1S, 0.2597; HP A*1V1, 0.0004; HP A*2FS, 0.7333; HP A*2SS, 0.0008; HP A*2V1, 0.0045; HP A*2V2, 0.0008; and HP A*2V3, 0.0004. The allele frequency of HP A*2SS, which is common in the European population, is less frequent than HP A*2V1 in the Japanese population.
Zymographic screening of plant peroxidase isoenzymes oxidizing 4‐hydroxystilbenes
Authors: Calderón Antonio A. , Pedreño M. Angeles , Ros‐Barceló Alfonso , Muñoz Romualdo 
Published: May 2005

AbstractA zymographic assay is described for the detection of peroxidase isoenzymes oxidizing 4‐hydroxystilbene following isoelectric focusing. The assay is based on coupling intermediate products of the oxidation of 4‐hydroxystilbene with 4‐aminoantipyrine, with resultant formation of dye complexes. Control experiments in the absence of 4‐hydroxystilbene and hydrogen peroxide demonstrate the peroxidative nature of the 4‐hydroxystilbene‐dependent dye reaction.
Microsequencing of proteins electrotransferred onto immobilizing matrices from polyacrylamide gel electrophoresis: Application to an insoluble protein
Authors: Hirano Hisashi , Watanabe Toshihiro 
Published: May 2005

AbstractSemidry electroblotting is convenient and allows a rapid and efficient protein transfer from one‐ or two‐dimensional polyacrylamide gels onto sequencer stable supports for protein microsequence analysis in a gas‐phase sequencer. Using this technique, we determined the amino acid sequences of the basic 7S globulin (Bg), an insoluble protein present in soybean seeds. Based on sequence determination, the cDNA‐encoding Bg could be easily cloned and characterized.
Calibration of polyacrylamide gel columns for the separation of oligonucleotides by capillary electrophoresis
Authors: Paulus Aran , Gassmann Ernst , Field Matthew J. 
Published: May 2005

AbstractPolyacrylamide‐filled gel columns are used to separate oligonucleotide samples. For homopolymeric standard samples, plots of migration time versus molecular size are presented over a range of 30–160 bases. With 2.5–4 % T and 3.3 % C gels, good resolution over the examined mass range, with peak width at half height of 3 to 6 s, is obtained by applying electrical fields of 200–400 V/cm. The separation of heteropolymeric nucleotides by slab gel electrophoresis under routine conditions was compared with capillary gel electrophoresis. Using the same column and the same separation conditions, the plot of migration time versus base number is linear with and identical slope for three oligonucleotide samples which were examined, allowing a calibration of a gel‐filled capillary for molecular mass determination.
Capillary zone electrophoresis separation and laser‐based detection of both fluorescein thiohydantoin and dimethylaminoazobenzene thiohydantoin derivatives of amino acids
Authors: Waldron Karen C. , Wu Shaole , Earle Colin W. , Harke Heather R. , Dovichi Norman J. 
Published: May 2005

AbstractCapillary zone electrophoresis is employed for the separation and analysis of both fluorescein thiohydantoin and dimethylaminoazobenzene thiohydantoin derivatives of amino acids. Detection of minute amounts of these amino acid derivatives is an important milestone in the development of a high sensitivity protein sequencer. Current detection limits for the fluorescein derivative is on the order of 10−21 moles whereas detection limits for the dimethylaminoazobenzene derivative is on the order of 10−16 moles.
Multiphasic electrophoresis with diversified spacers
Authors: Bringard Aline , Charlionet Roland 
Published: May 2005

AbstractThe principle and some applications of multiphasic electrophoresis with diversified spacers are described. The method combines previously published concepts of introducing highly diversified spacers into the stack of an isotachophoretic system with the theory of snow‐ and telescope‐electrophoresis. The link between this electrophoretic method and isotachophoresis on the one hand and monophasic zone electrophoresis on the other hand is exemplified. The importance of the nature of the spacers as well as of the pH and ions in the leading and terminating zone is illustrated. Details are given concerning the experimental set‐up. This electrophoretic technique could serve as an alternative to isoelectric focusing.
Isoelectric focusing in immobilized pH gradients of phosphoglucomutase and esterases from the spiny lobster
Authors: Espinosa Georgina , Wenisch Elisabeth , Danner Herbert , Katinger Hermann , Righetti Pier Giorgio 
Published: May 2005

AbstractA method is described for detecting polymorphisms of cephalothorax and tail homegenates of 25 puerulus staged Panulirus argus in phosphoglucomutase (PGM) a esterases. Isoelectric focusing in immobilized pH gradients was used. In the pH 6.0–8.0 interval for phosphoglucomutase and in the pH 3.5–5.0 and 4.2–4.9 rang for esterases both enzymes appeared as polymorphic band patterns. These could explained by one locus with 2 alleles for phosphoglucomutase and 3 loci with 2, 3 and 4 alleles for esterases. Esterases exhibit a more extensive polymorphism in immobilied pH gradients than in polyacrylamide gel electrophoresis.
Hereditary subtypic patterns detected in the Ba fragment of complement factor B: Occurrence of four common alleles in Japanese
Authors: Suzuki Koichi , Ito Shigenori , Tamura Akiyoshi , Fujita Kiyoshi , Matsumoto Hideo 
Published: May 2005

AbstractA procedure which can detect subtype‐specific minor bands of factor B (BF) by polyacrylamide gel isoelectric focusing is presented. After zymosan‐mediated fragmentation of BF in serum via alternative pathway for complement activation, serum samples are subjected to isoelectric focusing in a narrow pH range (4.2–4.9). The Ba fragments are detected by using immunoblotting. In addition to the previously reported minor bands with subtypic specificities, heterogeneities are observed in other minor band group, where a single minor band corresponds exclusively to a subtype in a regular combination with the previously announced subtypic patterns. A one‐to‐one correspondence of a single band to each subtype provides an unambiguous determination for three subtypic phenotypes deduced from the two divided BF*F alleles, BF*FA and BF*FB. An autosomal codominant heredity is confirmed through segregation analysis. A population survey reveals that four common alleles, BF*S, BF*FA, BF*FB, BF*Fb1, occur in a Japanese population and the former three alleles, except BF*Fb1, occur in a Cambodian population. The presence or absence of a single anodal minor band was found to be the only difference after neuraminidase treatment of FA and FB, implying that an amino acid substitution responsible for the FA–FB subtypic difference is involved in an additional acquisition in FA of an oligosaccharide unit with a charged sialic acid.
Analysis of caprine mycoplasmas and mycoplasma infections in goats using two‐dimensional electrophoresis and immunoblotting
Authors: Olsson Birgitta , Bölske Göran , Bergström Katrin , Johansson Karl‐Erik 
Published: May 2005

AbstractSeven stains of mycoplasma have been characterized by two‐dimensional polyacrylamide gel electrophoresis in combination with immunoblotting. The two‐dimensional patterns of the strains were compared and similarities and differences are discussed. Hyperimmune rabbit antisera were used in the immunoblotting experiment to study the immunological cross‐reactivity between the species. Sera from goats, naturally and experimentally infected with mycoplasma, were also used in the immunoblotting experiment to study the immune responses of the infected animals.
Two‐dimensional electrophoresis of the fatty acid binding protein from human heart: Evidence for a thiol group which can form an intermolecular disulfide bond
Authors: Nielsen Søren U. , Vorum Henrik , Spener Friedrich , Brodersen Rolf 
Published: May 2005

AbstractA 100 000 g supernatant from human heart muscle, containing cytosolic proteins with some contaminating plasma proteins, was analyzed for fatty acid binding protein (FABP) by two‐dimensional electrophoresis (2‐DE) using isoelectric focusing under nondenaturing conditions in the first dimension. FABP purified from human heart muscle was found to comigrate with a major spot in 2‐DE gels of the supernatant. This spot was comparable with those of the myoglobins in staining intensity. When purified FABP was charged with [H]palmitate and subjected to nondenaturing 2‐DE, radioactivity always comigrated with this protein. Under denaturing and reducing conditions in the second dimension, FABP was found to have a pI of 5.3 and an apparent molecular weight of 15 000. Isoforms of FABP, reported here for the first time to occur in human heart muscle, were observed as minor spots focusing at pH 5.1 and 5.7. When electrophoresis in the second dimension was carried out under denaturing but nonreducing conditions, an additional protein appeared at pH 5.3 with an apparent molecular weight of about 30 000. This protein was identified as a dimer of FABP and evidence for the involvement of an intermolecular disulfide bond in this dimerization is presented.
Electrophoresis of nonreduced IgM and IgG monoclonal antibodies on mixed agarose‐polyacrylamide gels
Authors: Slavíčková Alena 
Published: May 2005

AbstractA gel matrix composed of 0.5 % agarose and 2.5 % polyacrylamide allows the parallel electrophoretic separation of nonreduced IgM and IgG monoclonal antibodies. The mobilities of IgM and IgG differ at pH 9.5, which enables a preliminary Ig class designation. It allows rapid detection of incomplete cloning if both IgM and IgG are present. A semiquantitative evaluation of nonreduced IgM in ascitic fluid is possible as well.