Anion-exchange chromatography DEAE-cellulose

Anion Exchange of DEAE-Cellulose. When employed at a pH above the isoelectric point of the gonadotropins and at low ionic strength, it has been shown that DEAE-cellulose, used either batchwise or in column chromatography, adsorbs most of the FSH from a pituitary extract, and leaves most of the LH unabsorbed. 

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%. 

The, chain voAiantS are characterized by the presence of two abnormal components, an abnormal Hb-F (02 /2) and an abnormal Hb-A (tt2 32) Of these two, the 02 2 component dominates and the 02 32 component Is often difficult to detect. The methods of choice are starch gel electrophoresis and anion-exchange chromatography using DEAE-Sephadex or DE-52 Cellulose. Chain analyses of these Isolated hemoglobin components will lead to a definitive Identification. 

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.

Immunization and Fusion Protocol. Groups of five BALB/c female mice (4-6 weeks old), were given series of three injections every two weeks, with KLH-conjugated atrazine or hydroxyatrazine (50 ug/injection) mixed with Freund s adjuvant. After a rest period of two months, the mice were boosted with 500 ug of the conjugate. Three to four days later, the mice were sacrificed and the spleen cells were fused with the murine myeloma cell line Sp 2/0.Agl4 (12,13). The positive hybridomas were cloned and expanded in mice and the MAbs were purified from the ascitic fluid by ammonium sulfate precipitation, and DEAE-cellulose anion-exchange chromatography (14). 

Citric acid cycle acids Strong cation exchange resin acid elution no regeneration Anion exchange chromatography (beaded DEAE--cellulose) volatile ethanol-formic acid eluents Turkelson and Richards [104] Bruinsma and Le Tourneau [116]... 

After coupling, the acetyl groups are removed by brief treatment with methanolic ammonia. The product dinucleotides are purified by anion-exchange chromatography using DEAE-cellulose, and characterized spectroscopically. H-NMR spectra of the product carba-NAD is shown in Figs. 1 and 2. The spectrum of the dinucleotide exhibits the expected contributions from the pyridine nucleotide portion and from the adenosine nucleotide. 

Liver N-acetyltransferase can be partially purified by a procedure which includes ultracentrifugation (100,000 g), ammonium sulfate precipitation, gel filtration on Sephadex G-lOO and anion exchange chromatography on DEAE-cellulose resin (Weber and Cohen, 1967 Weber, 1971b). The total activity recovered in this procedure is about 15% of the activity in the 100,000 g supernatant fraction. A 300- to 500-fold purification from this fraction is achieved. Liver N-acetyltransferase from man, monkey (cynomolgous), rabbit, and rat has been purified according to this procedure. The enzyme from monkey (rhesus) liver has also been purified approximately 250-fold by a somewhat different procedure (Goedde et al., 1967). 

Figure 4. Anion-exchange chromatography on a DEAE-cellulose column of the P-III fraction from Sephadex G-200 gel chromatography and SDS-PAGE of the DEAE fractions (I-IV). The dotted peak (the DEAE-I fraction) was pooled. M, mol. wt markers.

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