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At least 19 recordsLinked to original sources

Size and shape of poly(ADP-ribose): examination by gel filtration, gel electrophoresis and electron microscopy.

Digestion of the poly(ADP-ribose) with snake venom phosphodiesterase yielded phosphoribosyl-AMP and AMP in a ratio of 30 to 1, indicating the "chain length" of 30 if the poly(ADP-ribose) is a linear molecule as previously supposed. Gel filtration and gel electrophoresis, however, revealed a very heterogeneous size distribution, the average size being, much longer than (ADP-ribose)30. Electron microscopy of poly(ADP-ribose) showed an irregularly branched figure. Molecules with more than ten branched portions were observed. These results are in agreement with the recent identification of the chemical structure of the branched portion in poly(ADP-ribose).

Animals↗

Polyethylene glycol interferes with protein molecular weight determinations by gel filtration.

Gel filtration studies on Sephadex G-75 demonstrate markedly increased elution volumes for proteins chromatographed with polyethylene glycol. As little as 5% (w/v) polyethylene glycol in the applied protein sample can reduce apparent molecular weight estimates by gel filtration as much as 55%. Furthermore, gel filtration columns equilibrated with polyethylene glycol are not size-separating columns. Consequently, caution must be exercised when performing and interpreting gel filtration studies of proteins previously treated or precipitated with polyethylene glycol.

Chromatography, Gel↗

Measurement of drug displacement of protein-bound bilirubin using gel filtration.

Gel filtration was used to measure drug interaction with protein-bound bilirubin in 0.5 ml samples of Gunn rat serum, human serum and fraction V human serum albumin. Using sulfadimethoxine as a prototype differences in displacement were found in all 3 sera. The differences between human and rat serum were related to the binding characteristics of sulfadimethoxine whereas the differences between human serum and fraction V human serum albumin were attributed to displacement of bilirubin from albumin to other proteins in serum. Gel filtration permitted the use of small samples with bilirubin-albumin ratios less than 1.0 and provided data that were used for analysis of drug displacement of bilirubin using principles of drug-receptor theory. Ten of 14 drugs found to alter serum bilirubin concentrations in icteric Gunn rats had measurable effects on protein binding of bilirubin.

Animals↗

Inoculation of BALB/c mice against Leishmania major infection with Leishmania-derived antigens isolated by gel filtration.

Gel filtration can be used as a substitute for preparative polyacrylamide gel electrophoresis in the preparation of an immunogenic, partly purified, fraction isolated from lysates of Leishmania promastigotes. This fraction, the molecular weight of which ranged from 70,000 to 53,000, when administered to BALB/c mice in associated with muramyl dipeptide induced resistance against cutaneous leishmaniasis.

Animals↗

Immunological properties of Friend virus from mouse spleen obtained by gel filtration.

Gel filtration, with Sephadex G-200, was utilized to obtain Friend virus (FV) from infected spleen homogenate. Highly infectious and immunologically active material was eluted as a large initial protein peak, whereas hemoglobin and other noninfectious materials were found in two subsequent peaks. Fractions that made up the first peak were reactive in serological tests with antiserum towards FV. Formalinized fractions from the first peak, when combined with Freund's complete adjuvant, were able to protect mice against subsequent FV challenge. Further purification of antigen was carried out by sucrose gradient ultracentrifugation.

Animals↗

Separation and quantitation of serum constituents associated with calcium by gel filtration.

Gel filtration of serum by use of polyacrylamide beads (Biogel P-2) separates total colcium into four distinct peaks: an initial peak, corresponding to protein-bound calcium; a second peak containing the calcium complexes of citrate, phosphate, lactate, and sulfate; a third peak containing calcium bicarbonate; and a fourth peak or trough corresponding to the serum ionic calcium. An eluent containing (per liter) 140 mmol of sodium, 1.1o mmol of calcium, 0.50 mmol of magnesium, and 10 mmol of 2-([tris(hydroxymethyl)methyl]amino)ethanesulfonic acid(pH 7.40 at 37 degrees C) provides physiological conditions that determine the equilibria between these calcium components. Association constants determined under these conditions permit calculation of the expected concentration of the calcium complexes in each tube of eluent, and these concentrations closely correspond to the amount of bound calcium measured experimentally. The mean distribution of calcium in healthy individuals, as determined by this method, is (per liter): calcium protein, 1.00 mmol; calcium complexes, 0.31 mmol; and ionic calcium, 1.07 mmol.

Bicarbonates↗

Preparative procedure for the purification of toxoids by gel filtration.

Sephadex gel filtration can be employed as a preparative procedure for the purification of both tetanus and diphtheria toxoids. A toxoid purification sequence is described in the text. By utilizing the described methods and columns, up to 100,000 human doses of diphtheria toxoid could be processed in a single operation. The method has given an 80% yield of diphtheria toxoid with a purity of 1,900 Lf per mg of N. The analysis of the material by immunodiffusion tests showed that a marked increase in purity was achieved. Antigenicity tests demonstrated that there was no significant difference in antigenic potency between the parent toxoid and its purified fraction. Factors limiting the effective separation of tetanus toxoid by gel filtration are discussed. The construction of the columns used is described in detail, as well as packing procedures and column characteristics such as bed volume, void volume, sample size, and flow rate.

Animals↗

Separation of oligogalacturonic acids by high-performance gel filtration chromatography on silica gel with diol radical.

Oligogalacturonic acids (OLGAs) ranging from two to nineteen residues in length were separated using high-performance gel filtration chromatography on a silica gel with diol radical. The optimum conditions (eluent, column temperature) for separation of OLGAs by high-performance gel filtration chromatography were investigated. The column used in this experiment allowed a high pressure of 4900 p.s.i. and a flow-rate of 2 ml/min. The stationary phase of silica gel stabilized the separation of OLGAs. The peaks of OLGAs separated using this column were assigned by comparing retention times with standards, and the molecular weights of the corresponding OLGAs were determined by fast atom bombardment mass spectrometry.

Chromatography, Gel↗

Phosphatases in lake water: characterization of enzymes from phytoplankton and zooplankton by gel filtration.

Sephadex gel filtration was used to characterize phosphomonoesterases in two small takes in northern Sweden. Two fractions, here termed phosphatase A and phosphatase B, were found both as free enzymes and associated with seston. The activity of phosphatase A was correlated with the presence of algal biomass. Phosphatase B, on the other hand, was derived from zooplankton. Phosphate served as an effective inhibitor of phosphatase A but had no such effect on phosphatase B. Both fractions had pH optima between 6.5 and 7.0.

Chlorophyta↗

Unified theory for gel electrophoresis and gel filtration.

Unified theory for gel electrophoresis and gel filtration: The behavior of macromolecules in gel filtration and gel electrophoresis may be predicted from Ogston's model for a random meshwork of fibers. This model has been generalized to apply to nonspherical molecules and to several gel types. The model provides equations for inter-relationships between mobility, partition coefficient, gel concentration, and molecular radius; it gives a non-Gaussian distribution of pore sizes as a function of gel concentration. The theory defines conditions for optimal separation and optimal resolution in gel filtration and gel electrophoresis. The difference in resolving power between the two fractionation methods is accounted for by the fact that gel filtration is a form of partition chromatography.

Chromatography, Gel↗