DEAE-cellulose ion-exchange

El 3. In what order would the following proteins be eluted from a DEAE-cellulose ion exchanger by an increasing salt gradient. The pHj is listed for each protein. 

DEAE-cellulose ion-exchange chromatography, which is dependent on the phosphate, pyrophosphate or phosphoethanolamine groups of the lipid A moiety, is widely used for lipid A isolation (El Hamidi et al., 2005 Raetz and Kennedy, 1973). TLC, as a lipids detection method, also can be applied for lipid A isolation (Zhou et al., 1999). In addition, the chromatography techniques based on the molecular... 

The sequential purification of T. borchgrevinki serum on a DEAE-cellulose ion-exchange column and a QAE-Sephadex A-25 column is seen in Figs. 4 and 5. Slab-gel electrophoretic patterns of the mixture of active AFGP components 1-5 and of the three main active... 

In 1965, Yonetani and Ray H) obtained a highly purified preparation of cytochrome c peroxidase in an excellent yield using DEAE-cellulose ion exchange chromatography. Shortly thereafter, Yonetani et al. 15) crystallized this enzyme by isoelectric dialysis. Subsequently, Yonetani and co-workers 16-40) carried out a series of extensive investigations... 

DEAE- Cellulose Ion exchange Amino acids 0.01-1 Maq. NaCI Ninhydrin ... 

DEAE cellulose, ion exchange, glucose oxidase and catalase. 

Pulich (4) had observed the autooxidation of low potential cytochrome c in extracts of Nostoc when supplied with NADPH under aerobic conditions and he suggested that cytochrome reduction was catalysed by ferredoxin-NADP oxidoreductase in his preparation. We could easily repeat Pulich s observation using purified proteins but found the rate of cytochrome reduction was very low. The addition of crude ferredoxin to the reaction mixture greatly stimulated the rate of electron transfer. When extracts of the soluble proteins of cyanobacteria are chromatographed on DEAE cellulose ion exchange columns, two forms of ferredoxin are occasionally observed. When cells are grown with sufficient iron, one finds a ferredoxin which elutes from the column al 0.4 to 0.5 M NaCl and which is usually identified with electron transfer from Photosystem I to FNR. This is called ferredoxin I. 

A proteinaceous inhibitor of rubber biosynthesis was purified from the C-serum of Hevea brasiliensis latex. The protein inhibited the incorporation of isopentenyl diphosphate into rubber. Purification was achieved by employing three column chromatography methods Sephadex G-150 gel filtration, DEAE-Cellulose ion exchange chromatography and Phenyl Sepharose CL-4B hydrophobic interaction chromatography. 21 refs. 

Fig. 10. Exchange reactions. A. Between AcCoA and H-CoA. The reaction mixture contained 0.01 ml of H-CoA solution (0.027 /imole), 0.07 ml of 4 X 10 M AcCoA (2.8 nioles) prepared in water, 0.10 ml of DEAE-cellulose enzyme fraction (30 lig protein) from a rapid inactivator human and 0.8M potassium borate buffer, pH 8.0 in a total volume of 0.27 ml at 27 °. Aliquots of 0.02 ml were removed and immediately mixed with 0.05 ml of 3 X 10" M DTNB dissolved in 0.05 M sodium acetate buffer, pH 5.0 (0.15 Mmoles) at 4° to stop the reactioa Aliquots of 0.01 ml were then removed from this mixture, and the Ac- H-CoA was separated from the H-CoA on DEAE-cellulose ion-exchange paper. The Ac- H-CoA area was cut out, placed in a counting vial, eluted with 0.02 ml of 0.6 N HCl and counted. Complete reaction mixture (o) mixture without AcCoA ( ) mixture with enzyme previously inactivated b y heating at 55° for 15 min (A). B. Between H-aniline and acetanilide. The reaction mixture contained 0.005 ml of H-aniline dissolved in water (115 mC/mmole, 0.0019 /tmole), 0.20 ml of 3 X lO M acetanilide dissolved in 0.1 M sodium pyrophosphate buffer, pH 8.0 (6.0 Mmoles), 0.10 ml of DEAE-cellulose enzyme fraction from a rapid inactivator human (30 Mg protein) and 0.10 M sodium pyrophosphate buffer, pH 8.0, in a total volume of 0.305 ml at 27°. Aliquots of 0.01 ml were removed, placed on CM-cellulose strips, and treated with 0.005 ml of acetone-absolute ethanol (1 1) to stop the reaction. The H-acetanilide was separated from the H-aniline by eluting the strips with glycine buffer at pH 2.8. The H-acetanilide area was cut out, placed in a count-...

Layers of cellulose ion-exchangers are prepared with the usual commercial spreaders. A mixture is made of water or the solvent to be used, and about 10—20% cellulose powder. The amount of powder depends on the swelling tendency of the particular powder. The exchangers CM-, DEAE- and ECTEOLA-ceUulose tend to swell considerably PEI- and Poly-P-cellulose only slightly. This swelling is disadvantageous in that the completely dry layer shows a greater or lesser number of very fine cracks. 

Ion exchangers can also be made from cellulose, especially for scientific applications. They are prepared from alkali cellulose by reaction, for example, with chloroacetic acid (for preparation of sodium carboxymethylcellulose, see Example 5-6). By conversion with 2-chloroethyldiethylamine one obtains so-called DEAE-cellulose, an anion exchanger carrying 2-diethylaminoethyl groups, -C2H4N(C2H5)2. 

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