Gel filtration chromatography columns

Gel-filtration column chromatography Determination of average molecular weight and the NH7-terminus... 

Fig. 4.5. Gel-filtration column chromatography of dansylated microbubble glycopeptide surfactant on Sephadex G-25. The column (0.9 cm I.D. x 55 cm) was equilibrated with 50 mM-Na acetate buffer, pH 4.5, containing 4 M guanidine-HCl. The column was eluted with the equilibrating buffer, at a flow rate of 3.3 ml per hour. The volume per fraction was 0.7 ml, and each fraction was combined with an added 0.8 ml aliquot of the equilibrating buffer before being analyzed for fluorescence. Activation of the dansyl label was performed at 254 nm, and the emitted fluorescent light was monitored at 450 nm. (Taken from ref. 322.)...

Gel-filtration column chromatography, amino acid analysis and carbohydrate determination... 

Further purification of the microbubble surfactant mixture by gel-filtration column chromatography... 

Several methods are available for determining the molecular weight of proteins. Gel-filtration column chromatography, SDS-PAGE, and ultracentrifugation are among the most commonly used. 

Figure 4-2. Size-exclusion chromatography. A A mixture of large molecules (diamonds) and small molecules (circles) are applied to the top of a gel filtration column. B Upon entering the column, the small molecules enter pores in the stationary phase matrix from which the large molecules are excluded. C As the mobile phase flows down the column, the large, excluded molecules flow with it while the small molecules, which are temporarily sheltered from the flow when inside the pores, lag farther and farther behind.

Remove excess crosslinker and reaction by-products by dialysis or size exclusion chromatography. For small quantities of bait proteins, dialysis may be the better choice, because gel filtration columns often bind nonspecifically enough protein to make recoveries unacceptably low. 

Retention of a protein or protein activity after 105,000y, 1 hr Chromatography on gel filtration columns with large pore sizes Electron microscopy—however, sample preparation may partially reconstitute membranes Decrease in solution turbidity, which may be detected by a diminution in light scattering or an enhancement in light transmission Diffusion of membrane lipids as assayed by nuclear magnetic resonance and electron spin resonance... 

A 10 mL round-bottomed flask was charged with (5)-6,6 -[oxybis(ethylene) dioxy]biphenyl-2,2 -diol (1) (98% ee, 225.6 mg, 0.78 mmol), quinine (127.0 mg, 0.39 mmol) and ethanol (2mL). The mixture was warmed until the mixture suspension turned to a clear solution, and was allowed to settle for 12 h. The solid residue [crystals of (5)-l-quinine complex] was collected by filtration. To a mixture of aqueous 1 M HCl and ether was added the obtained crystals of (5 )-l-qumine complex. This was stirred for 15 min at room temperature, and the solution was extracted with ether (twice). The combined organic layers were washed with brine, and dried over sodium sulfate. After concentration in vacuo, the residue was purified by silica gel flash column chromatography (hexane/ethyl acetate = 20/1-3/1) to give optically pure (5)-l (167.8 mg, 74%, 99% ee) as a colourless solid. The enantiomeric excess of (5)-l was determined by chiral stationary-phase HPLC analysis DAICEL CHIRALCEL... 

Experimental studies to construct a binding curve will be described at pH 4.5. To prepare the gel filtration column, obtain a chromatography column (about 1.5 X 15 cm). It should be equipped with a porous glass disk and a stopcock at the bottom to control flow rate. Clamp the column to a ring stand and connect the bottom tubing to a fraction collector containing 50 test tubes. If... 

Many schemes for fractionating nucleotides, nucleosides and bases on sulphonated polystyrene resins have been published. The main difficulty with these methods is variation between resin batches (e.g. Anderson et al. 1963). Nucleotide separations can be achieved on DEAE-cellulose (Whatman Data Sheet 13, 1967) and DEAE-Sephadex (Piers et al. 1965b) but these media do not seem to be widely used. Gel filtration columns will separate some nucleotide components. Ligand exchange chromatography and partition chromatography of nucleosides are useful for minor components. 

Figure 14.4. Selectivity curve from SEC of DNA ( ) and protein ( ) molecular weight standards on a 106 x 10 mm Superose 6 gel filtration column with 0.02 M Tris-HCl pH 7.6 containing 0.15 M NaCl as the eluent.4 [Reprinted, with permission, from H. Ellegren and T. Laas, Journal of Chromatography 467, 1989, 217-226. Size - Exclusion Chromatography of DNA Restriction Fragments. Fragment Length Determinations and a Comparison with the Behaviour of Proteins in Size-Exclusion Chromatography . 1989 Elsevier Science Publishers B.V.]...

After concentration, the solubilized enzyme was applied to a gel filtration column. The enzymes from pear and tomato were purified further (1.5-3 fold) by isoelectric focusing (20,25). Chloro-plastic hydroperoxide lyase was solubilized from tea leaves with Tween 20 and partially purified 8.5-fold with 34% recovery by hydroxyapatite column chromatography (33). Although attempts at... 

Janson and Hedman (1) recently published an excellent review of large-scale chromatography. Many of the broad process design and operation considerations are the same for affinity chromatography as they are for ion exchange or gel filtration. Most chromatography models, however, are based on the assumption of small feed pulses with linear equilibria (such as the widely-used plate theories (2)) and are not directly useful for affinity separations. In this paper we discuss and compare experimental results with two fixed-bed adsorption models that can be used to predict the performance of affinity columns. These two models differ only in the form of the rate-... 

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