Ammonium sulfate, extraction

Precipitate residual protein with hydrochloric acid and ammonium sulfate extract interfering lipids with methylene chloride derivatize aqueous fraction with pentafluorobenzaldehyde extract with ethyl acetate. 

Figure 1 shows the proteins contained in various subnuclear fractions. Histones were not detectable in the isolated nuclear matrix by Coomassie blue staining. But these known acceptor proteins for ADP-ribose were found in the ammonium sulfate extract. Autoradiography revealed that great amounts of the self-modified poly(ADP-ribose) synthetase - in accordance with the results of Table 1 - were released by DNase, RNase digestion of the isolated nuclei (not shown). 

Acetonitrile also is used as a catalyst and as an ingredient in transition-metal complex catalysts (35,36). There are many uses for it in the photographic industry and for the extraction and refining of copper and by-product ammonium sulfate (37—39). It also is used for dyeing textiles and in coating compositions (40,41). It is an effective stabilizer for chlorinated solvents, particularly in the presence of aluminum, and it has some appflcation in... 

The oxime is converted to caprolactam by Beckmann rearrangement neutralization with ammonia gives ca 1.8 kg ammonium sulfate per kilogram of caprolactam. Purification is by vacuum distillation. A no-sulfate, extraction process has been described, but incineration of the ammonium bisulfate recovers only sulfur values and it is not practiced commercially (14). 

Snia Viscosa. Catalytic air oxidation of toluene gives benzoic acid (qv) in ca 90% yield. The benzoic acid is hydrogenated over a palladium catalyst to cyclohexanecarboxyhc acid [98-89-5]. This is converted directiy to cmde caprolactam by nitrosation with nitrosylsulfuric acid, which is produced by conventional absorption of NO in oleum. Normally, the reaction mass is neutralized with ammonia to form 4 kg ammonium sulfate per kilogram of caprolactam (16). In a no-sulfate version of the process, the reaction mass is diluted with water and is extracted with an alkylphenol solvent. The aqueous phase is decomposed by thermal means for recovery of sulfur dioxide, which is recycled (17). The basic process chemistry is as follows ... 

Prior to the bating process, the hides are delimed with ammonium sulfate and/or ammonium chloride. Proteases are then appUed. The early preparation proposed by Rn hm was pancreatic trypsin. The use of a bating enzyme makes the hides soft and supple to prepare them for tanning. A new microbial protease, Pyrase 250 MP (82) (Novo Nordisk A/S) has been found to be a promising substitute for pancreatic trypsin [9002-07-7] which is more expensive because it must be extracted from pancreatic glands. 

The method of extraction of Ru(III) from thiocyanate solutions by water soluble extractants in the presence of ammonium sulfate as salting out agent followed by photometric determination of it in extract has been elaborated. 

A 500-ml, three-necked, round-bottom flask is fitted with a mechanical stirrer, a thermometer, and a wide-stern (powder) funnel. The flask is cooled in an ice-salt bath and charged with 125 ml (approx. 0.5 mole) of 15% sodium hydroxide solution. When the stirred solution reaches -10°, 30% hydrogen peroxide (57.5 g, 52.5 ml, approx. 0.5 mole) previously cooled to -10° is added in one portion. The pot temperature rises and is allowed to return to —10° whereupon 37.5 g (0.25 mole) of phthalic anhydride (pulverized) is added rapidly with vigorous stirring. Immediately upon dissolution of the anhydride, 125 ml (approx. 0.25 mole) of cooled (-10°) 20% sulfuric acid is added in one portion. (The time interval between dissolution of the anhydride and the addition of the cold sulfuric acid should be minimized.) The solution is filtered through Pyrex wool and extracted with ether (one 250-ml portion followed by three 125-ml portions). The combined ethereal extracts are washed three times with 75-ml portions of 40% aqueous ammonium sulfate and dried over 25 g of anhydrous sodium sulfate for 24 hours under refrigeration. 

The small jellyfish Phialidium gregarium (diameter 15-20 mm) used to be abundant at Friday Harbor, Washington, in summer and autumn until about 1990. Levine and Ward (1982) isolated and purified a Ca2+-sensitive photoprotein from this jellyfish and named it phialidin. They extracted the photoprotein from whole specimens with an EDTA-containing buffer. The photoprotein extract was precipitated with ammonium sulfate, and purified by the following methods gel-filtration (BioGel P-150, minus 400 mesh), anion-exchange chromatography (DEAE Bio-Gel A), and gel-filtration (Sephadex G-75, superfine). 

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... 

The frozen shells were ground in a cold mortar with 50 mM sodium acetate buffer, pH 5.8, containing 10 mM EGTA and 0.2 M NaCl, then the mixture was centrifuged. The pellets were re-extracted with the same buffer, and centrifuged. All supernatants were combined, and the photoprotein was precipitated with ammonium sulfate. The photoprotein in the precipitate was purified by four steps of column chromatography at near 0°C. Due to the instability of the photoprotein, efforts were made to reduce the time required for purification. 

Cormier and Dure (1963) found another type of luciferin and called it protein-free luciferin. Protein-free luciferin was found in the vapor condensate of freeze-drying whole animals, and also in the 3 5-56 % ammonium sulfate fraction of the crude extract noted above. The protein-free luciferin behaved like an aromatic or heterocyclic compound and it was strongly adsorbed onto Sephadex and other chromatography media, requiring a considerable amount of solvent to elute it. The luminescence reaction of protein-free luciferin in the presence of luciferase required a 500-times higher concentration of H2O2 compared with the standard luciferin preparation. Both types of the luciferin preparation had a strong odor of iodoform. 

By extraction from pineapple stems with water and precipitation with acetone or ammonium sulfate. Reference(s) ... 

A wheat germ, cell-free, translation extract was fractionated into three concentrated parts using ammonium sulfate the 0 - 40 % saturated fraction, the 40 - 60 % saturated fraction, and the ribosome fraction. These fractions were tested for their ability to enhance the translational activity of the wheat germ, cell-free extract for dihydrofolate reductase. The fortified cell-free system supplemented with the 0 - 40 % ammonium sulfate fraction enhanced the efficiency of protein synthesis by 50 %. 

Previously, it has been reported that the amounts of eukaryotic initiation factors in wheat germ extract prepared by a common method were deficient for the translation of some kinds of mRNAs including a-amylase mRNA and (i-globin mRNA [2]. Therefore, it can be expected that the activity of wheat germ extract prepared by a common method can be enhanced by the simple addition of extract containing deficient initiation factors. In this study, a wheat germ extract was further purified partially by ammonium sulfate fractionation... 

Fig. 1. Reconstruction of the cell-free protein synthesizing system with the partially purified wheat germ extracts. Control normal wheat germ cell-free system, (I) 0 - 40 % ammonium sulfate fraction 3 pi, 40 - 60 % ammonium sulfate fraction 4 pi, and ribosome 3 pi were added to 25 pi reaction mixture, (II) 0-40 % ammonium sulfate fraction 4 pi, 40 - 60 % ammonium sulfate fraction 4 pi, and ribosome 1.5 pi were added to 25 pi reaction mixture.

The catalytic activities of the fortified wheat germ cell-free systems supplemented with each fraction were investigated (Fig. 2). As shown in Fig. 2, only 0 - 40 % ammonium sulfate fraction showed an enhancement in DHFR protein synthesis. This enhancement of protein experimental results and the fact that the various eukaryotic initiation factors are contained in synthesis was also confirmed by SDS-PAGE and autoradiography (Fig. 3). From the above 0-40 % ammonium sulfate fraction [5, 6], it can be concluded that the amount of initiation factors in a conventionally prepared wheat germ cell-fi extract is deficient for the translation of DHFR with internal ribosome entry site. Therefore, it needs to supplement a wheat germ cell-free extract with the fraction containing the limited initiation factors for the efficient protein translation, and this fortified cell-free system can be easily made by simple... 

Ammonium Sulfate Precipitation. The extract was made up to 40% saturation with the slow addition, with stirring, of ammonium sulfate at 4°C. After several hours, the precipitate was removed by centrifugation at 30,000 g for 30 min and the supernatant retained. It was brought to 100% saturation in similar conditions, the precipitate was collected by centrifugation, dissolved in the minimum of distilled water, dialyzed against water and then against 1% glycine, and lyophilized. 

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