Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “ELECTROPHORESIS”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 757 records · Page 42Linked to original sources

STARCH GEL ELECTROPHORESIS OF A FRACTION OF CERTAIN OF THE PLEUROPNEUMONIA-LIKE GROUP OF MICROORGANISMS.

Fowler, Richard C. (George Washington University School of Medicine, Washington, D.C.), Don W. Coble, Norman C. Kramer, and Thomas McP. Brown. Starch gel electrophoresis of a fraction of certain of the pleuropneumonia-like group of microorganisms. J. Bacteriol. 86:1145-1151. 1963.-Starch gel electrophoresis is suggested as a means of further characterizing strains of the pleuropneumonia-like group of microorganisms (PPLO), defined herein as including both the various "L" forms of bacteria and the Mycoplasmataceae. The techniques used may be varied to "select" other groups of proteins or classes of compounds (e.g., glycoproteins, lipoproteins, and enzymes). Only the results by starch gel electrophoresis on the soluble portion of sonically treated, washed cultures, grown in a pancreatic digest of beef heart enriched with bovine serum, are reported. As yet, differences appear more significant than similarities among the electrophoretically separable proteins. The demonstration of characteristic protein patterns for each strain studied suggests possibilities of further dividing serological groups into sero-logically distinct subtypes. Tentative migration values (E(f)) have been assigned to the protein bands for convenience in designation. Numerous interrelationships are thereby suggested.

Animals↗

Improved measurement of monoclonal paraproteins in serum using agarose gel electrophoresis.

Serum paraproteins can be accurately and precisely measured by means of agarose gel electrophoresis and densitometry at 520 nm after staining with Coomassie Brilliant Blue G. The results obtained for this method agree closely with the quantity of paraprotein recoverable from preparative agarose gel electrophoresis. The method is preferable to cellulose acetate electrophoresis stained with Ponceau S.

Antibodies, Monoclonal↗

[Identification of monoclonal immunoglobulins by immunofixation electrophoresis on cellulose acetate films and in agarose gel].

A method of immunofixating electrophoresis is used for identification of monoclonic immunoglobulins, i.e. for estimation of heavy chain classes and of light chain types. Optimal conditions for immunofixation were developed. Advantages of immunofixating electrophoresis as compared with immunoelectrophoresis were established. Immunofixating electrophoresis on acetate cellulose films is recommended as a screening test for monoclonic immunoglobulins.

Electrophoresis↗

Comparison of electrophoresis on citrate agar, cellulose acetate, or starch for hemoglobin identification.

We compare and discuss three electrophoretic methods for identifying hemoglobins S, A, C, F, and D or G. Electrophoresis on citrate agar gel was more sensitive than electrophoresis on cellulose acetate for detecting hemoglobins S and F, a fundamental consideration in designing cord-blood screening programs for detecting hemoglobin S carriers. Electrophoresis on starch gel is evidently an acceptable method for subtyping hemoglobins AA, CC, AS, SS, AC, and SC, and is more sensitive than cellulose acetate for identifying hemoglobin A1A2. Costs for the citrate agar gel, cellulose acetate, and starch gel procedures are presented.

Adolescent↗

[Use of proteins, covalently bound to agar gel, for specific sorption in electrophoresis].

A method is developed for electrophoretic fractionation of serum proteins in agar gel with simultaneous specific sorption of gamma-globulin fraction, migrating during the electrophoresis through specific immunosorbents incorporated into agar gel. The reaction was carried out in a special apparatus designed for the electrophoresis in two perpendicular directions. The sorbent was separated from impurities by passing the current in the first direction. The unspecific sorbent did not alter the electrophoretic mobility of migrating proteins, but the specific sorbents immobilized completely both antibodies and antigens. The antibodies were shown to bind repeatedly to the immunoglobulins, which had been linked to the sorbent. The method facilitated distinctly the estimation of antigens and antibodies in complicated mixtures. By analogy with affinity chromatography a term "affinity electrophoresis" was used for designation of the method described.

Animals↗

[Polyacrylamide gel disc electrophoresis in the study of cerebrospinal fluid proteins in inflammatory diseases of the central nervous system Preliminary results].

The present study was designed to evaluate the potential of polyacrylamide disc electrophoresis of CSF proteins as an adjunct to laboratory diagnosis of neurological inflammatory diseases. The results of polyacrylamide and cellulose acetate electrophoresis of 42 CSF samples from control subjects and patients with various inflammatory diseases of CNS are presented. A comparison between results from both techniques is made. The polyacrylamide disc electrophoresis has been found valuable in the study of liquoral proteins with particular attention to gammaglobulins because this resolving power is superior to the other method. However, the Authors emphasize the importance of distinguishing the genetically determined proteins and haptoglobins and of the use of specific criteria in interpretation of a CSF protein pattern.

Cerebrospinal Fluid Proteins↗

Selective preconcentration for capillary zone electrophoresis using protein G immunoaffinity capillary chromatography.

Capillaries with 150 microns inner diameter were packed with a perfused protein G chromatographic support and used as immunoaffinity preconcentrators for capillary zone electrophoresis. Antibody was loaded onto the protein G support to form an immunoaffinity stationary phase. Injection of samples onto the column caused selective retention and preconcentration of antigen. Injection of appropriate buffers onto the column caused desorption of the antibody and antigen which were then separated by capillary zone electrophoresis. The combination was used on-line and off-line. For on-line combination, a flow-gated interface coupled the two columns and allowed injection of desorbed zones onto the electrophoresis system. Off-line coupling required collection of desorbed fractions and then injection onto the electrophoresis system. Flow rates as high as 100 microL/min were used to load sample onto the affinity column. Desorbing flow rates had to be 1 microL/min or less to prevent excessive dilution during desorption. Using the system, 1 mL insulin samples could be loaded onto the affinity column and desorbed in volumes as small as 1 microL for 1000-fold preconcentration. The use of the preconcentrator with serum samples spiked with insulin was demonstrated.

Affinity Labels↗

Molecular evolution among some Drosophila species groups as indicated by two-dimensional electrophoresis.

The evolutionary and phylogenetic relationships of seven Drosophila species groups (represented by D. melanogaster, D. mulleri, D. mercatorum, D. robusta, D. virilis, D. immigrans, D. funebris, and D. melanica) were investigated by the use of two-dimensional electrophoresis. The resulting phylogeny is congruent with the current views of evolution among these groups based on morphological characters and immunological distances. Previous studies indicated that the ability of one-dimensional electrophoresis to resolve relationships between distantly related taxa extended to about the Miocene [25 million years (Myr) ago], but the present study demonstrates that two-dimensional electrophoresis is a useful indicator of phylogeny even back to the Paleocene (65 Myr ago). In addition, two-dimensional electrophoresis is shown to be a useful technique for detecting slowly evolving structural proteins such as actins and tropomyosins.

Animals↗

Affinity titration curves: determination of dissociation constants of lectin-sugar complexes and of their pH-dependence by isoelectric focusing electrophoresis.

By performing electrophoresis perpendicular to a stationary pH gradient (pH-mobility curves) in polyacrylamide gels containing a specific ligand either covalently fixed or entrapped in the gel matrix, it is possible to measure dissociation constants (Kd) and their pH-dependence in the pH range 3.5-10. The present technique, called 'affinity titration curves', is an extension of 'affinity electrophoresis'. This system has been applied to the study of the interaction between lectins and sugars: lectin from Ricinus communis seeds and alpha-D-galactose, and lectin from Lens culinaris seeds and alpha-D-mannose. The pH-dependence of Kd values indicated a more rapid decrement of affinity of both lectins for their ligands at acidic pH as compared to alkaline pH. For both lectins, maximum affinity was found in the pH range 7-8. Since the ionic strength of focused carrier ampholytes is 100-200-times lower than in conventional electrophoresis, the Kd values found by the present method are generally lower than the same values obtained by affinity electrophoresis.

Carbohydrates↗

Application of monoclonal antibodies for affinity electrophoresis.

Monoclonal antibodies (MoAbs) to human alpha-fetoprotein (AFP) produced by 13 different murine hybridoma clones were used in affinity electrophoresis. The binding of MoAbs to AFP was studied in different experimental systems: crossed affinity immunoelectrophoresis, rocket affinity immunoelectrophoresis and zone affinity electrophoresis in agarose. Experimental data indicated that MoAb-AFP and AFP-MoAb-AFP complexes were formed during electrophoresis. Affinity electrophoresis seems to be a convenient and sensitive method for studies on the reaction of monoclonal antibodies with their antigens. In addition, the technique offers the possibility of calculating the dissociation constants of MoAb-AFP and of AFP-MoAb-AFP complexes and evaluating the reaction of MoAbs with different antigenic determinants.

Antibodies, Monoclonal↗

Limitations of differential electrophoresis for measuring colloidal forces: a Brownian dynamics study.

Differential electrophoresis experiments are often used to measure subpiconewton forces between two spheres of a heterodoublet. The experiments have been interpreted by solving the electrokinetic equations to obtain a simple Stokes law-type equation. However, for nanocolloids, the effects of Brownian motion alter the interpretation: (1) Brownian translation changes the rate of axial separation. (2) Brownian rotation reduces the alignment of the doublet with the applied electric field. (3) Particles can reaggregate by Brownian motion after they break, forming either heterodoublets or homodoublets, and because homodoublets cannot be broken by differential electrophoresis, this effectively terminates the experiment. We tackle points 1 and 2 using Brownian dynamics simulations (BDS) with electrophoresis as an external force, accounting for convective translation and rotation as well as Brownian translation and rotation. Our simulations identify the lower particle size limit of differential electrophoresis to be about 1 microm for desired statistical accuracy. Furthermore, our simulations predict that particles around 10 nm in size and at ambient conditions will break primarily by Brownian motion, with a negligible effect due to the electric field.

Journal Article↗

Free flow electrophoresis of chloroplasts.

Highly purified intact chloroplasts were isolated from spinach (Spinacia oleracea L.) leaves by free flow electrophoresis. Morphological and biochemical studies showed that the fraction enriched in intact chloroplasts has a higher protein to chlorophyll ratio and a higher linolenic acid content than the broken organelles of the other fraction. The intact chloroplasts prepared by electrophoresis retained their capacity for CO(2) fixation. Sodium dodecyl sulfate polyacrylamide gel electrophoresis demonstrated that this fraction was rich in stroma and lamellae proteins. Free flow electrophoresis, which separates organelles and molecules according to their surface charges, is a good technique for producing purified chloroplasts with complete physiological activities.

Journal Article↗

Plasma membrane vesicles of opposite sidedness from soybean hypocotyls by preparative free-flow electrophoresis.

Absolute orientations (sidedness) of plasma membrane vesicles obtained in highly purified fractions by preparative free-flow electrophoresis and by aqueous two-phase partition were determined based on ATPase latency and morphological criteria. Free-flow electrophoresis yielded two plasma membrane fractions. One, the least electronegative and designated fraction ;E,' was pure plasma membrane. The other, more electronegative and designated fraction ;C,' was heavily contaminated by various other cellular membranes. Plasma membrane vesicles from both fraction C and fraction E partitioned into the upper phase with aqueous two-phase partitioning. Purified plasma membrane obtained from microsomes by two-phase partition (upper phase) when subjected to free-flow electrophoresis also yielded two fractions, one fraction co-migrated with fraction C and another fraction co-migrated with fraction E. Both fractions exhibited an ATPase activity sensitive to vanadate and insensitive to nitrate and azide. ATPase activity was used as a structure-linked latency marker for the inner membrane surface. Concanavalin A binding (linked to peroxidase) was used as an imposed electron microscope marker for the outer membrane surface. Fraction E vesicles showed low ATPase latency (two-fold or less) and weak reactivity with concanavalin A peroxidase. In contrast, fraction C vesicles were characterized by much greater latencies upon detergent treatment (sevenfold) and a strong reaction with concanavalin A peroxidase. Two-phase partition as the initial procedure for plasma membrane isolation, yielded mixtures of vesicles of both inside out and right-side out orientation. Free-flow electrophoresis resolved the plasma membrane isolates into vesicles from fraction C which were right-side out (cytoplasmic side in), and vesicles from fraction E which were wrong-side out (cytoplasmic side out). Therefore, the two methods used in series, provided highly purified membrane preparations of apparently homogenous vesicles of opposite known absolute orientations.

Journal Article↗

Cell electrophoresis research directed toward clinical cytodiagnosis.

The application of cell electrophoresis to cytodiagnosis requires that a scientifically established basis exists for identifying abnormal cells electrophoretically, that research to detect such differences in the cytodiagnostic setting is possible and that a rapid and simple method of cell electrophoresis is adaptable to the clinical setting. Data are presented indicating modifications of electrophoretic mobility due to herpes simplex virus type 1 infection and Rous sarcoma virus transformation of culture cells. A simple apparatus for electrophoretically separating cells on a density gradient and collecting them for subsequent analysis is described, and results of experiments with this apparatus are consistent with those obtained by microscopic electrophoresis. Laser-doppler spectroscopic electrophoresis is suggested as a rapid method adaptable to clinical application.

Cell Line↗

Genetic typing of alpha1-antitrypsin by immunofixation electrophoresis, identification of subtypes of Pi M.

Prolonged electrophoresis in alkaline agarose gels, followed by immunofixation, is a valuable addition to acid starch gel electrophoresis and isoelectric focusing for the genetic phenotyping of alpha1-antitrypsin. This technique is helpful in clarifying certain variants and in ascertaining types in serum and amniotic fluid samples with secondary changes. In addition, heterogeneity may be detected within the variants found at pH 4.95, analogous to hemoglobin polymorphism. Two "new" variants, PiMLamb and PiMBaldwin, have been detected by a combination of immunofixation electrophoresis and acid starch gel electrophoresis.

Genetic Techniques↗

[Effectiveness of treating chronic cholecystitis by prodigiozain electrophoresis].

136 patients suffering from chronic cholecystitis were treated in the phase of exacerbation with antibiotics in combination with prodigiosan. It was found that inclusion of prodigiosan into the therapeutic complex had a positive effect on the immediate and late results of the treatment. Since intramuscular administration of prodigiosan was accompanied by a number of side effects, a method of the drug administration by means of electrophoresis on the area of the bile bladder was tested. It was shown that prodigiosan administered by the method of electrophoresis totally preserved its stimulating capacity with respect to the host. As with intramuscular administration of prodigiosan, the general reaction of the host and positive dynamics of the clinical and laboratory indices of the active process were observed and the content of lysozyme in the blood phagocytes reliably increased. The method of prodigiosan administration by means of electrophoresis was better tolerated by the patients and no side effects characteristic of the drug intramuscular administration were noted. When indicated, the use of prodigiosan by means of electrophoresis is recommended.

Adult↗

Separation of X- and Y-chromosome-bearing murine sperm by free-flow electrophoresis: evaluation of separation using PCR.

The effectiveness of separation of murine X- and Y-bearing sperm by free-flow electrophoresis was evaluated by the polymerase chain reaction (PCR). The ratio of X- and Y-bearing sperm from cauda epididymis was analyzed before and after free-flow electrophoresis. A Y-chromosome-specific sequence (pY353/B) and an autosomal sequence (myogenin) were used to estimate the ratio between X- and Y-sperm in the separated fractions. Cauda epididymal mice sperm were separated into two peak fractions under the electric field. Each peak fraction contained sperm of normal shape, however, the motility of the sperm was extremely diminished after separation by electrophoresis. DNA was extracted from 10(4) sperm from each fraction and from the unseparated sperm, and Y-chromosome specific PCR was performed. The PCR experiment revealed that fraction No. 16 (the peak near the cathode) was a Y-sperm rich fraction, whereas fraction No. 22 (the peak near the anode) was a Y-sperm poor one. These results suggested that murine X- and Y-sperm could be successfully separated by free-flow electrophoresis. Analysis of the chromosome-specific sequence by PCR was demonstrated to be a direct and adequate method to evaluate the separation of X- and Y-sperm.

Animals↗

[Interstitial electrophoresis in acute pleural and lung suppuration].

Different variants of interstitial electrophoresis elaborated in the hospital were used with the aim of raising the effectiveness of treatment of acute abscess of the lungs, of abscessing pneumonia and acute empyema of the chest. The principle of electroelimination of medicinal preparations from the vessels or directly from the suppurative cavity into the surrounding tissue is the basis of interstitial electrophoresis. As the result of experimental and clinical observations the following variants of interstitial electrophoresis are proposed: intrapulmonary, endobronchial, cavitary, intracavitary and combined. The use of the proposed method of treatment in acute pleuropulmonary suppuration permitted to achieve substantial raising of the effectiveness of the used complex therapy. The method of interstitial electrophoresis is recommended for wide use in practical pulmonary and thoracic surgery.

Acute Disease↗