DEAE cellulose chromatography

When the purified enzyme is subjected to ion-exchange chromatography (DEAE-cellulose) or gel filtration (Sephadex G-150), the three activities elute together with constant ratio in all fractions. 

Abras precatorius. Purified by successive chromatography on DEAE-Sephadex A-50, carboxymethylcellulose, and DEAE-cellulose. [Wei et al. J Biol Chem 249 3061 1974.]... 

Purified by Sephadex G-200 filtration and DEAE-cellulose column chromatography. Hexosaminidase A was further purified by DEAE-cellulose column chromatography, followed by an ECTEOLA-cellulose column, Sephadex-200 filtration, electrofocusing and Sephadex G-200 filtration. Hexosaminidase B was purified by a CM-cellulose column, electrofocusing and Sephadex G-200 filtration. [Srivastava et al. 7 Biol Chem 249 2034 1974.]... 

Alkaline phosphatase from rat osteosarcoma has been purified by acetone pptn, followed by chromatography on DEAE-cellulose, Sephacryl S-200, and hydroxylapatite. [Nair et al. Arch Biochem Biophys 254 18 1987.]... 

Reverse transcriptase (from avian or murine RNA tumour viruses) [9068-38-6] [EC 2.7.7.49]. Purified by solubilising the virus with non-ionic detergent. Lysed virions were adsorbed on DEAE-cellulose or DEAE-Sephadex columns and the enzyme eluted with a salt gradient, then chromatographed on a phosphocellulose column and enzyme activity eluted in a salt gradient. Purified from other viral proteins by affinity chromatography on a pyran-Sepharose column. [Verna Biochim Biophys Acta 473 1 7977 Smith Methods Enzymol 65 560 1980 see commercial catalogues for other transcriptases.]... 

Thrombin (from bovine blood plasma) [9002-04-4] Mj 32,600 [EC 3.4.4.13]. Purified by chromatography on a DEAE-cellulose column, while eluting with O.IM NaCl, pH 7.0, followed by chromatography on Sephadex G-200. Final preparation was free from plasminogen and plasmin. [Yin and Wessler J Biol Chem 243 112 796S.]... 

DEAE-cellulose chromatography. The 50% ethanol solution is poured onto a column of DEAE cellulose (2.6 x 10 cm), and F adsorbed at the top is eluted with 20 mM Tris buffer, pH 7.5, containing 0.2 M NaCl. A low pressure of argon gas is applied to accelerate the flow rate. The fractions containing F are combined, concentrated, and desalted using ethanol. 

Dried shrimp was ground, defatted with benzene, and then extracted with cold water. The luciferase extracted was purified first by a batch adsorption onto DEAE cellulose (elution with 0.4 M NaCl), followed by gel filtration on a column of Sephadex G-150, anion-exchange chromatography on a column of DEAE-cellulose (gradient elution 0.05-0.5 M NaCl), and gel filtration on a column of Ultrogel AcA 34. The specific activity of the purified luciferase was 1.7 x 1015 photons s 1 mg-1, and the yield in terms of luciferase activity was about 28%. 

Purification of aequorin. The purification method of aequorin reported by Shimomura et al. (1962) was essentially the repetition of column chromatography on DEAE-cellulose, the only usable, efficient chromatographic adsorbent available at the time. Since then, various different types of chromatographic media have been developed, and the purification method has been steadily improved. 

Trainor (1979) modified the above method (1) In the initial extraction, luciferin was extracted with 50 mM acetate buffer (pH 4.75) at 95°C, instead of boiling 20% methanol, to increase the extraction yield. (2) In the DEAE-cellulose chromatography, the column, on which the luciferin sample had been adsorbed, was washed with the following solvents before the elution of luciferin water, lOmM HC1 in methanol, methanol, and NaCl-saturated methanol. (3) To eliminate salts in purified luciferin, the solution was evaporated to dryness, and the luciferin in the residue was extracted with... 

Purification of luciferin (Rudie etal., 1976). The luciferin fractions from the DEAE-cellulose chromatography of luciferase were combined and concentrated in a freeze-dryer. The concentrated solution was saturated with ammonium sulfate, and extracted with methyl acetate. The methyl acetate layer was dried with anhydrous sodium sulfate, concentrated to a small volume, then applied on a column of silica gel (2 x 18 cm). The luciferin adsorbed on the column was eluted with methyl acetate. Peak fractions of luciferin were combined, flash evaporated, and the residue was extracted with methanol. The methanol extract was concentrated (1 ml), then chromatographed on a column of SephadexLH-20 (2 x 80 cm) usingmethanol asthe solvent. The luciferin fractions eluted were combined and flash evaporated. The residue was... 

Chromatography of hemoglobins on columns of DEAE-Cellulose (DE-52 mlcrogranular, preswollen Whatman) often results In excellent separations of many variants, because the hemoglobins are eluted as sharp, narrow zones generally widely separated from each other The hemoglobin zones are eluted from the DE-52 columns at a distinctly higher pH value than from a similar DEAE-Sephadex column ... 

Figure 2. Anion-exchange chromatography on DEAE Cellulose of fraction IP obtained after size-exclusion separation. Fractions (2 ml each) were pooled as indicated, o - Uronic acids, A - pentoses, x - hexoses.

Figure 2. CM-cellulose chromatography of pectolytic enzymes. The activity peaks of the flow-through of a DEAE-cellulose chromatography was applied to a CM-cellulose column. The column was eluted with a NaCl (0-0.5M) continuous gradient at a flow rate of 34 ml/h. 10 ml fractions were collected and assayed for pectolytic activities Symbols (0) pectate lyase ( ) polygalacturonase (reducing sugar-releasing activity) (x) protein. Other details in Methods.

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