Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Polyacrylamides”

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 631 records · Page 35Linked to original sources

[Penicillin amidase from E. coli. Some physicochemical properties of the enzyme incorporated into polyacrylamide gel].

The physico-chemical properties of penicillinamidase (PA) immobilized in polyacrylamide gel (IPA) were investigated. It was shown that simple incorporation of PA into polyacrylamide gel was not effective because of gradual washing out of the enzyme. The use of a complex method for the immobilization (immobilization in the presence of a linking agent) resulted in higher stability of IPA, the choice of the optimal ratio of the reagents being of paramount importance. The mechanical strength of IPA was studied in model experiments.

Acrylamides↗

A simplified method of protein electroblotting after isoelectric focusing on an ultrathin polyacrylamide gel layer fixed on a cellophane support.

A method is described for isoelectric focusing of proteins, using an ultrathin-layer polyacrylamide gel on cellophane, followed by electrophoretic transfer of separated proteins onto a nitrocellulose membrane. The polyacrylamide gel is firmly attached to the cellophane and thus protected from mechanical damage; such gels are easily manipulated. Cellophane is permeable to ions and application of this gel support overcomes difficulties resulting from the removal of ultrathin gels from a plastic support on electroblotting. Proteins separated under nondenaturing conditions were transferred onto a nitrocellulose membrane and detected by the concanavalin A-peroxidase technique. The proposed approach makes it possible to analyze the variability of nondenatured proteins and glycoproteins of different origin.

Acrylic Resins↗

Purification of anti-apolipoprotein CIII antibodies by exploiting their affinity for apolipoprotein CIII linked to polyacrylamide gel after isoelectric focusing.

A new immunoaffinity technique for purifying anti-apolipoprotein CIII antibodies has been developed by isolating bands of apo CIII2 from isoelectric focusing (IEF) gel. Total apo C was obtained from delipidated very-low-density lipoprotein by chromatography on Sephacryl S 200. Apo CIII2 was separated from the apo CII band by IEF on polyacrylamide gel with use of pH 4-6 ampholytes. After sonication of the band in distilled water and various different washes, we directly mixed with antiserum a suspension of apo CIII2 bound to IEF-polyacrylamide gel. After their elution, we tested the specificity of the antibodies by an enzyme immunoassay technique, using plates coated with apolipoprotein, 100 ng per well. No cross-contamination of eluted anti-apo CIII antibodies by anti-apo CII or anti-apo E antibodies was observed. This affinity technique is easy to use, rapid to perform, and no sophisticated apparatus is needed. The gels can be used repeatedly and yield reproducible results with a very good analytical recovery (94%). We anticipate that this technique will prove useful for purification of other antibodies, particularly antibodies to apolipoproteins such as anti-apo CII and anti-apo E.

Acrylic Resins↗

[Selection of Escherichia coli and Pseudomonas putida cultures with an enhanced resistance to immobilization on polyacrylamide gel].

Clones of Escherichia coli (A4, A70, G60) and Pseudomonas putida (A70, G30) with an elevated resistance to the process of immobilization in polyacrylamide gel and to the action of monomeric acrylamide were selected from the parent E. coli IBPM B115 and P. putida. The isolated cultures remained resistant to the above actions for a long time. The frequency at which cells with the elevated resistance appeared was comparable with the frequency of bacterial mutations. The plasmid analysis did not reveal the presence of plasmid DNA in the cells of the isolated cultures. The decrease in the viability index of bacterial populations caused by their immobilization in polyacrylamide gel and by the action of monomeric acrylamide did depend on the growth phase. The cells were more resistant to these actions in the stationary phase. The isolated cultures were more resistant as compared to the parent cultures irrespective of the growth phase.

Acrylamides↗

[Identification of multiple molecular forms of lactate dehydrogenase during isoelectric focusing with ampholines in polyacrylamide gel thin layer].

Multiple molecular forms of lactate dehydrogenase fractionated by isoelectric focusing with ampholines in the polyacrylamide gel thin layer were identified in the supernatant fluid of the myocardium and liver homogenate of the intact rats and rabbits. Differential detection of the enzyme fractions was based on differences in their substrate dependences and sensitivity to specific inhibitors. Simultaneously the analysis was made for the partially purified preparations of lactate dehydrogenase five isoenzymes from the same sources obtained by means of the preparative electrophoresis in polyacrylamide gel and ion-exchange adsorption on DEAE-Sephadex A-50. The isoelectric points of the identified fractions are determined. It is shown that redistribution of the enzyme multiple molecular forms within the pH gradient is subordinated to a certain strictly regularity: fractions from LDH1 to LDH5 occupy successively the sites in a weak-acid, neutral and weak-alkaline regions of the pH gradient.

Animals↗

Specific adhesion of rat hepatocytes to beta-galactosides linked to polyacrylamide gels.

Rat hepatocytes, isolated by a collagenase perfusion technique, specifically bind to polyacrylamide gel containing covalently immobilized 6-aminohexyl beta-D-galactopyranosyl groups. Less than 5% of these cells bind to polyacrylamide or to gels with the following covalently linked ligands: 6-aminohexanol, or the 6-aminohexyl D-pyranosides of alpha-mannose, beta-glucose, beta-2-acetamido-2-deoxyglucose, beta-cellobiose, beta-maltose, or beta-melibiose. Cell binding to beta-D-galactoside gels occurs after a lag period at 37 degrees and 65 to 100% (depending on the cell preparation) of the cells adhere. The duration of the lag period is inversely related to the beta-D-galactoside content of the gel but preincubation of the cells at 37 degrees reduces the lag period. Cell-gel binding is a threshold phenomenon. Adhesion of cells to gels does not occur when the glycoside concentration is less than about 900 nmol per cm2 x 0.25 mm thick gel piece. Above this critical concentration, cell-gel binding occurs and becomes maximal when the concentration is increased by only 20%. If these in vitro results apply to cellular interactions in vivo, they suggest that slight changes in the levels of cell surface or extracellular matrix carbohydrates may profoundly influence the behavior of neighboring cells.

Acrylamides↗

[Polyacrylamide gel disc electrophoretic study of laboratory and freshly isolated strains of Bordetella pertussis and Bordetella parapertussis].

Differences in the antigenic complexes of laboratory and newly isolated B. pertussis and B. parapertussis strains have established by means of disc electrophoresis in polyacrylamide gel. B. pertussis strains have been found to contain 2 or 3 antigens more. Laboratory and newly isolated B. pertussis strains have been found to differ in antigenic complexes, the antigens in these complexes having different electrophoretic mobility. The antigens detected in newly isolated strains have greater electrophoretic mobility in comparison with that of the antigens detected in laboratory strains. No such differences have been revealed in laboratory and newly isolated B. parapertussis strains. Disc electrophoresis in polyacrylamide gel allows to detect 2 or 3 more antigens than immunoelectrophoresis in agar gel.

Antigens, Bacterial↗

Adhesion of Dictyostelium discoideum cells to carbohydrates immobilized in polyacrylamide gels. II. Effect of D-glucoside derivatives on development.

In an accompanying report (Bozzaro, S., and Roseman, S. (1983) J. Biol. Chem. 258, 13882-13889), evidence is presented that the slime mold Dictyostelium discoideum contains three cell surface receptors specific for D-glucose, D-mannose, and N-acetyl-D-glucosamine, respectively. The synthetic probes used for these studies consisted of the sugars covalently linked to polyacrylamide gels. In the present experiments, starved cells were placed on these and other immobilized sugars to determine whether the sugar derivatives influenced normal development in this organism. When D. discoideum cells are on a solid surface under water, they form aggregation centers and strands of cells (which radiate from the center), send "signals" i.e. pulses of cyclic AMP from the center down the strands, and finally, after cells in the strands migrate to the center, form tight aggregates. These results were obtained on all polyacrylamide gel derivatives tested except one class, derivatives of D-glucose (O- and S-glucosides, cellobiosides, and maltosides). On these gels, aggregation centers and strands formed normally, but at a certain point stopped "signaling" and suddenly dissociated, with the cells rapidly migrating away from one another by negative chemotaxis (see Appendix to this report). Furthermore, a simultaneous dissociation of several centers was often observed. Following a brief period of random movement after dissociation, aggregation centers once again formed and the cycle was repeated. This cycle was repeated as often as 30 times or more over a 24-h period. The cells on the glucoside gels became aggregation-competent at the same time as the control cells, and the adhesion-dissociation cycle appeared to have no effect on the synthesis of some developmentally regulated proteins, such as UDP-glucose pyrophosphorylase. Interpretations of the phenomenon and its potential for studying gene regulation in this organism are discussed.

Acetylglucosamine↗

Free thyroxine in serum as estimated by polyacrylamide gel filtration.

In this new method for estimating free thyroxine concentration in serum by use of polyacrylamide gel filtration, the prolonged incubation that is characteristic of equilibrium dialysis is obviated. Although the values so obtained for the percentage of free thyroxine are severalfold higher than those obtained by dialysis, the results of the two procedures correlate well over a wide range of thyroid states. In addition, the precision of the polyacrylamide method compares well with that for equilibrium dialysis. The method is convenient for use in clinical laboratories and values for free thyroxine in serum of patients with metabolic thyroid disorders are diagnostically discriminatory.

Acrylamides↗

[Prediction of microorganism resistance to the immobilization process in polyacrylamide gel].

It is shown that the immobilization of bacterial cells in polyacrylamide gel or their exposure to monomer acrylamide results in a quantitatively similar decrease of their viability. It is indicated that acrylamide treatment may be used as a test for measuring the resistance of microbial populations to polyacrylamide gel immobilization and predicting the survival rate of microorganisms incorporated.

Acrylic Resins↗

System for simplified discontinuous-gradient polyacrylamide-gel electrophoresis.

We describe a modified polyacrylamide gel electrophoresis method with which plasma proteins are excellently resolved in 4 h. We made a three-layer discontinuous gradient gel with concentrations of 80,60, and 45 g of polyacrylamide per liter of tris(hydroxymethyl)aminomethane/ethylenediaminetetraacetic acid/borate buffer, pH 9.2. Time and material needed in the preparation of the gel was greatly decreased, a well-resolved (22-25 bands) pattern was obtained in 1 h, and the gel slabs were easier to remove from the cell than were gels prepared according to the manufacturer's specifications. We saw a marked shift in the higher polymers of haptoglobin type 2-2 toward the cathode in this system, and also a clearly resolved double band of alpha2-macroglobulin in whichthe more anodal band appears to represent alpha2-macroglobulin/protease complex.

Blood Protein Electrophoresis↗

Aberrant interaction of B-cell lymphocytic leukemia cells with antibody coated polyacrylamide beads.

Anti-immunoglobulin coated polyacrylamide beads have been used to assess surface membrane immunoglobulin in normal and lymphoproliferative states. The polyvalent anti-immunoglobulin bead reacts with normal B cells and cells from patients with hairy cell leukemia and lymphosarcoma cell leukemia but not with cells from patients with chronic lymphocytic leukemia. This may relate to the relative surface immunoglobulin density of these cells. Monospecific beads and mixtures of monospecific beads do react with chronic lymphocytic leukemia cells presumably because of the higher concentration of the appropriate antibody on the bead. These findings suggest cautious interpretation of the results of B lymphocyte evaluation with anti-immunoglobulin coated polyacrylamide beads.

Antibodies, Anti-Idiotypic↗

[Polarization fluorescence of chloroplasts oriented in polyacrylamide gel. Experiment and theoretical analysis].

Maize mesophyll chloroplasts were oriented in polyacrylamide gel upon squeezing of gel samples. Fluorescence polarization was measured as a function of the gel deformation parameter. Linearly polarized fluorescence spectra were recorded at -140 degrees. The results show that chloroplasts were oriented in polyacrylamide gel as effectively as in magnetic field. Theoretical models are analyzed for orientation of disc-shaped and rod-shaped chloroplasts. Of these two, only the model for disc-shaped chloroplasts can account for the experimental results. Based on experimental data, the approaches are proposed to estimate the order of alignment of thylakoid membranes within chloroplasts, and to calculate the angles made by transition dipoles with the thylakoid membrane plane. Internal structure of chloroplasts (alignment of thylakoid membranes) is shown to be unchanged upon squeezing of the gel samples.

Acrylic Resins↗

[Preparation of monospecific anti-Salmonella LPS antibodies from bacteria immobilized in polyacrylamide gel].

The use of an immunoabsorbent obtained by entrapping Salmonella cells into a polyacrylamide gel lattice enabled us to obtain anti-O monospecific immune sera which can be used for a quick serological identification of some species of Salmonella in the course of a diagnosis. In this paper we describe a method for immobilization of S. typhimurium as well as the preparation of monospecific anti-05 antibodies from plurispecific anti-S. haifa rabbit immune sera. This separation of anti-O monospecific antibodies avoids the repeated and often tedious adsorption of anti-Salmonella immune sera by the corresponding whole bacteria. Such Salmonella, entrapped into the polyacrylamide gel lattice, can be used several times without appreciable loss of their affinity properties.

Acrylic Resins↗

Investigation of the antigenic composition of the cells and the exotoxin of Sh. dysenteriae (Shiga) by agar immunoelectrophoresis and polyacrylamide gel disc electrophoresis.

Using the method of immunoelectrophoresis in agar, we established the presence of 12 soluble antigens in Sh. dysenteriae (Shiga). We differentiated among them 3 specific antigens situated in the cathodic region. One of them is the somatic O antigen while the other two are thermolabile surface K antigens of these bacteria. K antigens of Grigoryev-Shiga bacteria differ from the earlier described K antigens of the Newcastle and Boyd Shigellae in their positive electric charge, in greater lability in relation to temperature and chemical effects and evidently also in their chemical character. The remaining protein antigens are common for either the genus of Shigella or for the family of enterobacteria. The exotoxin preparation isolated from the R strain of Sh. dysenteriae (Voile 30) by precipitation with trichloroacetic acid is represented in the immunophoreogram by five precipitation lines. Disc electrophoresis in polyacrylamide gel made possible the investigation of the composition of the exotoxin and isolation of the two fractions responsible for its toxic effect. The method of preparative electrophoresis in polyacrylamide gel can evidently be used for obtaining purified Sh. dysenteriae exotoxin preparations.

Animals↗

[Properties of immunoadsorbents prepared by antigen coupling to glutaraldehydeactivated polyacrylamide gel, BrCN-activated Sepharose and by copolymerization of antigens by glutaraldehyde].

Three types of immunoadsorbents were synthetized by antigen coupling to glutaraldehyde-activated polyacrylamide gel, BrCN-activated sepharose 4B and protein insolubilization using glutaraldehyde as a cross-linking agent. Different concentrations of rabbit gamma globulin, bovine serum albumin, soybean trypsin inhibitor and bovine ribonuclease were used as antigens and some properties of such immunoadsorbents were studied. Using 125J labelled antigens it was shown that glutaraldehyde-activated polyacrylamide gel (Bio-Gel P-300) couples 81-94% of the antigen added in a concentration of 0.5-4.0 mg.ml-1 gel and BrCN-activated sepharose 4B bounds 62-88% of the labelled antigen in a concentration of 5.0-20.0 mg.ml-1 gel. These antigen derivatives as well as those obtained by glutaraldehyde protein insolubilization permitted 54-88% of antibodies added with the immune sera to be isolated. There was a significant antigen leakage from sepharose immunoadsorbents after several antisera treatments or after half a year storage but despite of this antigen desorption all types of the immunoadsorbents studied preserved their antibody isolation capacity. Immune sera immunoglobulins nonspecific binding by immunoadsorbents was less on sepharose than Bio-Gel antigen derivatives.

Antigen-Antibody Complex↗

Dry polyacrylamide batch method for studying drug protein binding.

The validity of a new technique for studying drug protein binding is tested. By this gel diffusion or batch method using dry polyacrylamide, sulfadiazine and sulfafurazole are bound by human plasma proteins to the same extent as reported by other authors with different techniques. The binding of warfarin to fresh human plasma was quantitatively less than expected. This might be due to the possibility that other plasma constituents of non-fasted test persons displace warfarin from protein binding sites, since it is bound more in buffer solutions containing physiological concentrations of human albumin. Phenylbutazone displaced warfarin from its binding sites in diluted human serum albumin concentrations. Quinidine was found to be adsorbed by the otherwise inert polyacrylamide gel and the method is therefore not applicable to quinidine binding studies.

Chromatography, Gel↗

Electrophoresis in microfabricated devices using photopolymerized polyacrylamide gels and electrode-defined sample injection.

Microfabrication techniques have become increasingly popular in the development of the next generation of DNA analysis systems. While significant progress has been reported by many researchers, complete microfabricated integrated DNA analysis devices are still in the earliest stages of development. Most miniaturized analysis systems have incorporated noncross-linked polymer solutions as the separation medium of choice and the operation of these systems necessitates the use of high electric fields and long separation lengths. In this paper, we present two techniques that may help alleviate this problem and accelerate the development of the so-called 'lab-on-a-chip' systems. We present the use of photodefinable polyacrylamide gels as a sieving medium for DNA electrophoresis. These gels offer the significant advantages of faster curing times, locally controlled gel interface, and simpler handling over chemically polymerized gels. We also introduce an electrode-defined sample compaction and injection technique. This technique helps achieve sample compaction without migration into the gel and offers significant control over the size and application of the sample plug. The use of these technologies for double-stranded DNA separations in microfabricated separation systems is demonstrated.

Acrylic Resins↗