bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel method for permanent imprints from dye‐stained electrophoretograms
Authors: Ramasamy Muthiah , Wimmer Dave H. , Kurup Viswanath P. 
Published: May 2005

AbstractA simple method for obtaining permanent imprints of Coomassie Blue and Amido Black stained electrophoretograms on developed unexposed photographic paper and film is described. Copies of slab and two‐dimensional electrophoretograms and crossed immunoelectrophoretograms were obtained by this technique. The method is fast, simple, inexpensive and can be adapted to make as many as five copies by repeated staining of the electrophoretogram. The copies are the exact size of the original gels and can be stored for a long time. The possible use of these imprints for the comparative study of immunoblots with the corresponding protein‐stained gels is discussed.
Non‐destructive sequential spread tissue isoelectric focusing
Authors: D'Andrea Alberto L. , Torres Leedhamsp VeróNica M. 
Published: May 2005

AbstractA method of sequential spread tissue isoelectric focusing is described, based on spreading a small amount of defrosted tissue, from musclebiopsy, on a rectangular filter‐paper applicator. A single tissue sample is applied sequentially at six different sites of an agarose gel and after focusing for contact intervals, between gel and applicator, of 15, 5, 5, 5, 5 and 15 min, isoelectric focusing is continued until 1500 Volt × hours are reached. The six patterns observed for a single tissue sample were qualitatively similar and sections of the same gel could thus be used for different detection methods, such as protein stainig, enzyme visualization or immunological detection.
Microscale electrophoresis of stress proteins induced by chemicals during the in vivo cell cycle
Authors: Pipkin James L. , Anson Jeanne F. , Hinson William G. , Robert Burns E. , Casciano Daniel A. 
Published: May 2005

AbstractThe incorporation of leucine and phosphate in proteins extracted with two different solvents from nuclei sorted physically from the G0+G1 phase of the rat submaxillary gland cell cycle was observed by two‐dimensional microscale polyacrylamide gel electrophoresis. Sodium phenobarbital did not initiate additional isotope ([3H]leucine and [32P]phosphate) incorporation on its own. After dosing with DL‐isoproterenol, several proteins incorporated leucine and phosphate during the G0+G1 phase. The inclusion of phenobarbital in the dosing regime enhanced labeling of several proteins already present in the G0+G1 phase. Treatment of animals with sodium arsenite prompted labeling of phosphate of two “stress” proteins in the non‐dividing G0 phase of the submaxillary gland. Enhanced incorporation of proteins was exhibited after combined dosing of isoproterenol and sodium arsenite, but no new incorporated proteins were seen in the G0+G1 phase. Likewise, no new proteins were seen to incorporate in the G2+M phase, but there was some increased incorporation in a few already present proteins under these conditions. The similar mobilities of several stress proteins, which were determined by coelectrophoresis of stress marker proteins, suggested that several of these proteins, even though they were extracted with different solvents, were identical.
Multiple forms of serum aminopeptidases separated by micro two‐dimensional electrophoresis under non‐denaturing conditions
Authors: Sanderink Ger‐Jan C. M. , Artur Yves , Galteau Marie‐Madeleine , Wellman‐Bednawska Maria , Siest Gérard 
Published: May 2005

AbstractMicro two‐dimensional electrophoresis in the absence of denaturing agents allowed the analysis of the multiple forms of serum aminopeptidases, including their multimolecular complexes, and the detection of their enzymatic activity towards different substrates. Two forms of alanine aminopeptidase (E.C. 3.4.11.2.), differing in sialic acid content, were detected in sera of healthy subjects. In sera of patients with hepatobiliary diseases a large number of additional spots was observed, some of them identified as multimolecular complexes, probably formed by an amphiphilic form of alanine aminopeptidase. Papain treatment resulted in the dissociation of the complexes and in the appearance of a light form of the enzyme in normal sera, which was otherwise only present in pathological samples. Additional spots corresponding to cysteine aminopeptidase (E.C. 3.4.11.3.) were detected in pregnancy sera.
Protein staining by Coomassie Brilliant Blue G‐250: Unique color rendering in iron‐toned prints
Authors: Sippel Theodore O. 
Published: May 2005

AbstractBlack‐and‐white photographs of protein staining by Coomassie Brilliant Blue G‐250 may be iron‐toned by a modified procedure to obtain good reproduction of the original hue and range of intensities. A suitable paper and a film for transparencies are specified for printing from a given negative.
Computer simulation of two electrophoretic columns coupled for isoelectric focusing in simple buffers
Authors: Tsai Amos , Mosher Richard A. , Bier Milan 
Published: May 2005

AbstractComputer simulation is used to analyze a system of two electrophoretic columns coupled by mixing the anolyte of one with the catholyte of the other. A mathematical model is presented which is used to predict the pH gradients formed by monovalent buffers in this system, when the currents in the columns are unequal. In the column with the higher current a pH gradient is created which increases from anode to cathode and is potentially useful for isoelectric focusing. The breadth of this gradient is dependent upon the ratio of the currents. The function of the second column is the compensation of buffer migration which occurs in the first column, thereby maintaining constant electrolyte composition. The effects of buffer pKs and mobilities are evaluated.
High resolution two‐dimensional electrophoresis of hair proteins without prior S‐carboxymethylation
Authors: Marshall Thomas , Williams Katherine M. 
Published: May 2005

AbstractHair proteins have been successfully analysed by high resolution two‐dimensional electrophoresis by avoiding S‐carboxymethylation which has previously dictated the use of alternative two‐dimensional procedures. The high and low‐sulfur proteins are simultaneously resolved within the pI range 3–9 and their electrophoretically determined Mr values are consistent with molecular weights obtained by physical methods.
The use of polyacrylamide gels that can be dissassembled and reassembled to permit a wide variety of electrophoretic procedures with a common polymerization history
Authors: Horowitz Paul , Bowman Steven 
Published: May 2005

AbstractA demountable polyacrylamide gel system is described that uses commercial components and can be adapted to any slab system. The use of GeldBond PAG film produces polyacrylamide gels that can be easily handled and allows separation of the conditions needed for polymerization from those for electrophoresis. Polymerization is performed with one common protocol, and the gel is disassembled. Gels can then be soaked or stored in any desired solutions, followed by reassembly of the equilibrated gel and electrophoresis. This procedure allows for a wide range of electrophoretic conditions starting with a single type of polymerized gel as well as permitting use of systems that have technical difficulties under normal cirumstances such as horizontal gels or those containing the cationic detergent, cetyltrimethylammonium bromide.
Application of recycling isoelectric focusing for purification of recombinant human leukocyte interferons
Authors: Nagabhushan Tattanahalli L. , Sharma Basant , Trotta Paul P. 
Published: May 2005

AbstractA computer‐controlled version of recycling isoelectric focusing has been applied to the purification of recombinant human leukocyte interferons (IFNs) from E. coli. The system is based on continuous recycling between a 12‐channel heat‐exchange reservoir and a 12‐channel focusing cell. It contains ten pH meters with probes in each of the flowing channels and two ultraviolet spectrophotometers with five flow cells per instrument. A minicomputer system acquires real‐time data of pH and optical density measurements, controls and continuously monitors voltage and power outputs across the focusing cell, and generates on‐line multi‐color graphics on an x‐y plotter. Recycling isoelectric focusing in the presence of 4 M urea resulted in removal of contaminating bacterial pyrogens from recombinant IFN alpha‐2 with complete retention of specific antiviral activity. It also produced a 25‐fold purification of crude recombinant IFN alpha‐5 in a single step and resolved two biologically active species. These data suggest that recycling isoelectric focusing may be incorporated into the design of simple schemes for the purification of a variety of recombinant proteins.
Affinophoresis of anti‐hapten antibody in rabbit serum with an anionic affinophore bearing the hapten
Authors: Shimura Kiyohito , Kasai Ken‐ichi 
Published: May 2005

AbstractAffinophoresis is an electrophoretic separation technique for biomolecules using an affinophore, which is a macromolecular polyelectrolyte bearing affinity ligands. It migrates rapidly in an electric field, and consequently the electrophoretic mobility of molecules having affinity for the ligand is specifically changed. An anionic affinophore bearing an antihypertensivepeptide, N‐(dibenzyloxyphosphinoyl)‐L‐alanyl‐L‐prolyl‐L‐proline, was synthesized by coupling the peptide to about one‐sixth of the amino groups of poly‐L‐lysine with an average degree of polymerization of 190. The remaining amino groups were succinylated and aminomethanesulfonic acid was coupled to about one‐fourth of the succinyl groups by the formation of amide bonds. Electrophoresis of rabbit antiser a raised against the peptide was carried out in an agarosegel plate containing the affinophore. After electrophoresis, separated proteins were transferred to a nitrocellulose sheet and immunoglobulins were visualized by using horseradish peroxidase‐conjugated anti‐rabbit immunoglobulin G antibody. Hapten‐specific antibodies were separated from non‐specific antibodies. Two‐dimensional agarose gel electrophoresis, in which affinophoresis was used in the second dimension, separated the hapten‐specific antibodies from other serum proteins.
A simplification of Heukeshoven and Dernick's silver staining of proteins
Authors: Damerval Catherine , Guilloux Martine Le , Blaisonneau Joël , de Vienne Dominique 
Published: May 2005

AbstractThe procedure of Heukeshoven and Dernick for silver staining of proteins in polyacrylamide gels is simplified by applying Farmer's redücer before impregnation with silver nitrate and rinsing the gel in 2.5 % sodium carbonate instead of distilled water. This modification yields pattern of good contrast without a destaining‐restaining step.
Separation of soybean leghemoglobin components by isoelectric focusing in immobilized pH gradients
Authors: Puppo Alain , Rigaud Jean 
Published: May 2005

AbstractSoybean leghemoglobin subcomponents were separated with good resolution in immobilized pH gradients, both at the analytical and preparative scales. In the latter case, the tedious step of gel washing could be omitted without affecting resolution. This procedure appeared as the only one suitable for preparing large amounts of highly purified leghemoglobin c1, c2 and c3, allowing their further characterization and an investigation of their specific roles in nitrogen fixation.
Human transcobalamin isopeptides: Separation by isoelectric focusing and by polyacrylamide gel electrophoresis
Authors: Hansen Mads , Fràter‐Schröder Marijke 
Published: May 2005

AbstractTranscobalamin was separated by isoelectric focusing and by polyacrylamide gel electrophoresis. Isoelectric focusing showed one band in homozygotes and two bands in heterozygotes, with isoelectric points between 6.3 and 7.0, while polyacrylamide gel electrophoresis produced two and four bands, respectively. Addition of neuraminidase, urea or EDTA had no effect on the separation of transcobalamin at isoelectric focusing. The results obtained by the two methods are comparable and in accord with the nomenclature of transcobalamin variants.
Determination of cell mixtures by an automated cell electrophoretic instrument and monoclonal antibody
Authors: Hayashi Haruhisa , Toyama Naomi , Fujii Masahiko , Yoshikumi Chikao , Kawai Yoshio , Iwaguchi Takao 
Published: May 2005

AbstractAn automated cell electrophoretic instrument, Parmoquant‐L, is based on the opto‐electronic image analysis whose principle is the same as conventional cytopherometers. Experiments are performed with high precision (C.V. values < 1.5 %) and in short time intervals of 5 min. Using this instrument the cell populations of a mixtureof sheep red blood cells (SRBC) and rabbit red blood cells (RRBC) were determined. The ratio of high mobility cells (HMC) to low mobility cells (LMC) was identical with the mixed ratio of SRBC to RRBC (R = 0.998) and there was no influence of different cell diameters between the two populations. As the electrophoretic mobilities of mouse red blood cells (MRBC) and SRBC were almost the same, the mixture of the two populations showed a broad unimodal electrophoretic mobility pattern. After the mixture was incubated with anti‐SRBC monoclonal antibody (diluted to 1/100), the mobility of SRBC decreased 30 % and it was possible to determine the cell numbers of each erythrocyte in the mixture. From these results, Parmoquant‐L is suitable to determine the populations in the mixture of different erythrocytes.
Two‐dimensional electrophoresis with immobilized pH gradients in the first dimension: Protein focusing as a function of time
Authors: Hanash Samir M. , Strahler John R. , Somerlot Luke , Postel Wilhelm , Görg Angelika 
Published: May 2005

AbstractSince the introduction of immobilized pH gradient (IPG) electrophoresis, optimization of sample entry has been a major concern. Partial sample entry, associated with smearing of proteins, has led investigators to propose the use of hybrid IPG‐carrier ampholyte focusing for adequate protein separation. We have examined the effects of adding carrier ampholytes to the protein sample, preelectrophoresis and the site of sample application on sample entry, resolution and pattern stability for IPG gels, used as a part of two‐dimensional electrophoresis. The addition of carrier ampholytes to the sample increased the rate of entry of proteins into the gel while preelectrophoresis had an opposite effect. There was a substantial difference in protein entry between sample application at the anode vs. cathode. After an adequate focusing time the final focusing pattern was remarkably similar and consisted of several hundred well‐resolved polypeptides, irrespective of the initial starting conditions. Therefore IPG electrophoresis, without addition of carrier ampholytes to the gel, results in the high resolution separation of complex protein mixtures.
A modified applicator strip for improved focusing on immobilized pH gradient gels
Authors: Pflug Werner , Laczko Bela 
Published: May 2005

AbstractThe use of applicator strips on isoelectric focusing in immobilized pH gradient gels often results in lateral sample spreading and mixing of adjacent samples. Both effects can be reduced or eliminated if a modified applicator strip with slits at the bottom is used. Different modes of sample application were compared using the group specific component system with different concentrations of bovine serum albumin in the samples.
Reduced area two‐dimensional gels for direct digital imaging of radioactive protein profiles
Authors: Ross Mary E. , Latter Gerald , Burbeck Stephen , Leavitt John 
Published: May 2005

AbstractA method is demonstrated which reduces the area of a standard two‐dimensional polyacrylamide electrophoresis gel to approximately one‐fourth of its original area. Microdensitometry is used to show that the technique did not significantly alter the reproducibility of the gels. This reduction in size makes it possible to use a micro‐channel plate analyzer to directly digitize an entire gel.
Moving boundary electrophoresis on agarose gel of plant viruses and polystyrene microspheres
Authors: Gombocz Erich , Tietz Dietmar , Chrambach Andreas 
Published: May 2005

AbstractThree plant viruses: turnip crinkle (TCV), hibiscus chlorotic ringspot (HCRSV) and pelargonium flowerbreak (PFBV), and polystyrene size standards with radii of 22.4–59.4 nm can be stacked within trailing and leading ion net mobilities of 0.059 to 0.273 (relative to Na+). Stacking was carried out at pH 6.50, 0.03 M ionic strength, 50 mM 3–[(3‐cholamidopropyl)‐dimethylammonio]‐1‐propane sulfonate, at a gel concentration of 0.4 %, in agarose gel electrophoresis conducted at 1.2 mA/cm2 of gel. Unstacking occurs under the same conditions at gel concentrations ranging from 0.5 to 1.1 % agarose, while it can be brought about between 0.1 and 0.7 % agarose when the pH is raised to 7.27, corresponding to a front moving boundary with a trailing ion net mobility of 0.216 (relative to Na+). Ferguson plots of viruses and polystyrene particles in the discontinuous buffer system are curvilinear and comparable to those obtained in a continuous buffer at pH 6.50 of the same composition and operative pH as that of the resolving phase of the discontinuous buffer. Particle radii and net charge values can be obtained from the non‐linear Ferguson plot in the discontinuous buffer system by previously reported methods of computer simulation, but this Ferguson plot presents a more limited data base than that in the continuous buffer since it excludes gel concentrations which yield relative mobility (Rf) values of 1.0. Since computer simulation provides the range of gel concentrations in which both the fiber radius and length [1], as well as the size of the particle [2], remain constant for a particular preparation of agarose, a simplified alternative method of evaluating particle sizes exists. Within that specific gel concentration range, the linear segment of the Ferguson plot can be used to compute particle sizes by an operationally convenient, albeit approximative, method, using the same PAGE‐PACK programs of D. Rodbard which are commonly used in the size determination of macromolecules by polyacrylamide gel electrophoresis. The two methods of particle size determination, based on either the entire non‐linear Ferguson plot or on its linear segment in the appropriate gel concentration range, yield similar results (average deviation 12 %). The radii of three plant viruses are dependent to different degrees on the presence of Ca++ in the electrophoretic system. Values obtained in the presence of Ca++ are comparable to those found by electron microscopy.
Affinity electrophoresis with monoclonal antibodies
Authors: Breborowicz Jan , Gan Jacek , Klosin Jadwiga , Gryska Katarzyna , Majewski Przemyslaw , Breborowicz Danuta , Waliszewski Przemyslaw 
Published: May 2005

AbstractAffinity electrophoresis was applied to the study of binding of monoclonal antibodies (MoAb's) with their respective antigens. The MoAb's may be either mixed with the antigen in an application well or incorporated into the agarose gel. The first approach is more convenient for studying high affinity MoAb's, while MoAb's with low affinity might be more efficiently studied in the second system. Our conditions for affinity electrophoresis do not imply nor necessitate immobilization of either an antigen or MoAb and are most suitable for studies of MoAb's reacting with antigens of alpha mobility demonstrated in experiments with MoAb's against human alpha‐fetoprotein (AFP). Both AFP‐MoAb and AFP‐MoAb‐AFP complexes were observed during electrophoresis. The identity of these complexes was confirmed by their reaction both with polyclonal antibodies against mouse immunoglobulins and against human AFP. The molecular weight of complexes was determined by gel filtration. Affinity electrophoresis was less convenient for studies of MoAb's reacting with antigens of gamma mobility at pH 8.6. Nevertheless, MoAb's against carcinomembryonic antigen, human IgG and human IgA did bind with their respective antigens under the conditions of affinity electrophoresis. The binding of MoAb's with these antigens during affinity electrophoresis was confirmed by demonstration of mouse immunoglobulins in the precipitates formed by reaction of the studied antigens with polyclonal antibodies. Our results point out the suitability of affinity electrophoresis for studies of MoAb's.
Fast preparative isoelectric focusing of phycocyanin subunits in layers of granulated gels
Authors: Köst‐Reyes Eliana , Schneider Siegfried , John Wolfhart , Fischer Richard , Scheer Hugo , Köst Hans‐Peter 
Published: May 2005

AbstractA new method is presented for the fast preparative separation of the light‐harvesting photosynthetic pigment C‐phycocanin into its α and β subunits, which is based on isoelectric focusing in layers of granutaled gels containing 7 M urea. The method has been successful in cases where other separation procedures failed. The recovery of the separated chains of the light ‐sensitive biliprotein amounts to 70 ± 10 % when the separation is carried out under light exclusion and in an argon atmosphere. A simple and inexpensive setup for work under an atmosphere of protective gas is described.
The effect of prestaining prior to immunoreaction during electrophoretic blotting of proteins
Authors: Tracy Russell P. , Monkovic Don , Andrianorivo Aurelie , Calore James , Mackay Alastair 
Published: May 2005

AbstractWe have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson, Electrophoresis, 1984, 5, 35–42) to blotting of unstained gels with Amido Black or Fast Green post‐transfer staining of the nitrocellulose. Contrary to a recent report (Harper et al., Anal. Biochem., 1986, 157, 270–274), in which post‐staining with Coomassie Blue was determined to severely inhibit immunoreactivity, we find using prestained Coomassie Blue gels to be extremely useful in most cases, although, rarely, inhibition of the immune reaction may take place. Post‐staining with Amido Black or Fast Green may also prove useful, but is not recommended since similar inhibition occurs, and there are the added disadvantages of not being able to “pre‐view” the gel pattern and not being able to use stored gels. Further studies indicated that the rare loss of immunoreactivity seen with Coomassie Blue prestained gels is most likely due to the necessary fixation step, and not the stain itself (as has been suggested), since the protein‐dye complex is dissociated during transfer.
Simplified molding procedure for ultrathin‐layer polyacrylamide slab gels using methacrylate supports and unsilanized glass plates: Hybrid isoelectric focusing on ultrathin immobilized pH gradients
Authors: Pascali Vincenzo L. , Destro‐Bisol Giovanni , d'Aloja Ernesto 
Published: May 2005

AbstractA simplified procedure for the preparation of ultrathin polyacrylamide slab gels is reported. The molding cells consist of an unsilanized glass plate and methacrylate plate or acryl‐coated glass plate. Owing to the repelling properties of acrylate the ultrathin gels spontaneously detach from the acrylate surface on unfastening the molds, firmly adhering to the unsilanized glass. Gels as thin as 0.20–0.25 mm could be polymerized without resorting to silanized glass plates or polyester films. The procedure is especially suitable for sticky gels, such as highly cross‐linked polyacrylamide gels and gels with immobilized pH gradients.