Dialyzer reactions

Dialyzer reactions encompass a broad range of clinical symptoms that include anaphylactic (type A) and nonspecific (type B) events. In the past, these two types of reactions were considered to be part of the first-use syndrome because they occurred much more frequently when new, as opposed to reprocessed, dialyzers were used. Although reprocessing may reduce the incidence of type B events, it has little to no benefit for patients who experience a type A reaction. The... 

In one manufacturing process, aluminum chloride is treated with a solution containing sodium carbonate and sodium bicarbonate. The product of this reaction is mixed with the precipitate obtained by reaction of a solution of aluminum chloride and ammonia. The mixed magma is dialyzed, the product mixed with glycerol (qv), sodium benzoate is added, and the mixture is then passed through a coUoid mill. 

A mixture of l,4-dibromo-2,5-bis(3-sulfonatopropoxy)benzene 61 (0.78 g, 1.39 mmol), 60 (0.23 g, 1.39 mmol), Na2C03 (0.99 g) in doubly distilled water (47 mL), and DMF (20 mL) was heated at 85°C until the solids were completely dissolved. The resulting solution was cannulated to a 200-mL Schlenk flask with tris[(sulfonatophenyl)phosphine]palladium(0) (0.045 g) and the mixture was stined at 85°C for 10 h. The reaction mixture was concentrated to 25 mL by boiling and filtered. The filtrate was added dropwise to cold acetone (250 mL) to precipitate out the polymer. The polymer was collected by filtration, redissolved in a minimum of hot water, and reprecipitated by cooling. After repeating this procedure twice, the polymer was redissolved in distilled water and dialyzed for 72 h in 3500 gmol 1 cutoff membrane. After drying under vacuum, polymer 63 was obtained in 64% (0.42 g). 

To prepare the mild-alkali-extract, dry watermelon cell walls were suspended in a solution of 0.1 N NaOH, and allowed to react with stirring at room temperature for 15 minutes. A pH of 13, as indicated by pH paper, was kept constant during this period by addition of 0.1 N NaOH. To ensure complete reaction, the treatment was continued overnight at 4 °C. The soluble portion was separated by centrifugation at 10,000 RPM for 20 minutes in a Sorval GSA rotor. The insoluble portion was washed twice with water. The supernatants were combined and, after neutralization to pH 7.0 with acetic acid, dialyzed against distilled water and freeze dried. 

The enzyme had a requirement for calcium. The addition of EDTA to the reaction mixtures, resulted in complete loss of activity, whereas the addition of CaCl2 increased the activity (figure 8). Presumably, sufficient contaminating calcium ions were present in the dialyzed enzyme and substrate mixture to permit the limited activity of the controls, but apparently these were removed by chelation with EDTA. The optimum concentration was in the range of 5 to 15 M, and higher concentration resulted in a decrease in activity. Phoma medicaginis var. pinodella synthesizes a pectin lyase that lacked an absolute requirement for calcium ions but maximum enzyme activity required the presence of 1 mM Ca [25]. The lyase from Fusarium solani f sp. phaseoli, that is active on pectin and pectic acid, is calcium-dependent [30]. Most of the pectate lyases characterized are calcium-dependent the pectate lyase from Rhizoctonia solani [34] and the endopectate lyase fi om Fusarium solani f sp. pisi [31]. 

The reaction vessel is then packed in ice and the reaction mixture is neutralized to pH 6.5 to 7.0 with aqueous hydrochloric acid. The neutral solution was dialyzed against nitrogen-saturated, distilled water for two days. The polymer solution is precipitated in a 5- to... 

The water-soluble hydroxyethylcellulose has only a limited use as compared to water-soluble methylcellulose, principally because of the difficulty of isolating the product. Aqueous solutions of hydroxyethylcellulose do not gel on heating and the product is about as soluble in hot water as in cold, so that the method used for isolating water-soluble methylcellulose is not applicable. However, by dialyzing the reaction product the alkali and salts may be removed and the product isolated. 

To remove excess TCEP and reaction by-products, dialyze the solution or purify by gel filtration using a buffer containing 1-10 mM EDTA. 

Label a set of tubes according to the standards and samples to be used. Add 250 pi of each standard and sample to the appropriate tubes. If the samples are in a buffer that may significantly change the pH of the reaction buffer, the samples should be buffer-exchanged or dialyzed into 0.1M sodium phosphate, pH 8.0, before running the assay. 

Extensively dialyze the reaction mixture against water or buffer to remove excess reactants. 

Immediately purify the maleimide-activated protein by applying the reaction mixture to a desalting column. Do not dialyze the solution, since the maleimide activity will be lost over... 

Dialyze the oxidized liposomes against 20mM sodium borate, 0.15M NaCl, pH 8.4, to remove unreacted periodate. This buffer is ideal for the subsequent coupling reaction. Chromatographic purification using a column of Sephadex G-50 also can be done. 

Dialyze against PBS, pH 7.2 to remove excess reagent and reaction by-products. 

To remove excess CDI and reaction by-products, Beauchamp et al. (1983) dialyzed against water at 4°C. However, the imidazole carbamate groups on mPEG formed during the activation process are subject to hydrolysis in aqueous environments. A better method may be to precipitate the activated mPEG with diethyl ether as in the protocol described previously for SC activation (Section 1.2, this chapter). 

Dissolve the bait protein to be modified with the FeBABE reagent in 50 mM MOPS, 100 mM NaCl, ImM EDTA, 5 percent glycerol, pH 8.2 (coupling buffer). The protein first may be treated by dialysis with EDTA to remove residual metals (30mM MOPS, 4mM EDTA, pH 8.2), and then dialyzed into the final coupling buffer for the reaction with FeBABE. 

The addition of more pancreatic amylase to reaction mixtures that had been dialyzed during the hydrolysis and that had reached stages of... 

Amine-terminated, G3 (PAMAM) dendrimer, (0.316 g 45.7 moles) was dissolved in anhydrous methyl sulfoxide (5 ml) in a 100 ml round-bottom flask flushed with dry nitrogen. After dendrimer had completely dissolved, succinic anhydride (Aldrich) (0.363 g 3.6 mmol) was added to the reaction mixture with vigorous stirring, and the mixture was allowed to react for 24 h at room temperature. The product solution was diluted with deionized water, transferred to 3500 MWCO dialysis tubing (Spectrum) and dialyzed against deionized water (18 Mil) for 3 d. The retentate solution was clarified by filtration through Whatman No. 1 filter paper, concentrated with a rotary evaporator, and lyophilized to yield a colorless powder (0.435 g, 94%). The product was analyzed by 13C-NMR, FT-IR, SEC and MALDI-MS. The analytical data were consistent with the expected carboxylic acid-terminated product. 

DCM-Dex was synthesized according to Scheme 2. Dextran (500 mg) was dissolved in 8.8 M NaOH (10 mL). Bromodiethylmalonate (10 g) dissolved in 10 mL of tetrahydrofuran was added to the solution at 0°C. The solution was then stirred at room temperature for 20 h. The reaction mixture was dialyzed in distilled water using cellulose tubing for 7 days. 

We have examined whether the sulfur that was bound to the proteins of a reconstituted system from the liver of phenobarbital-treated rats was bound to both the reductase and cytochrome P-450. I this experiment, the reconstitued system was incubated with [ s] parathion. The reaction mixture was dialyzed and applied to a Sephadex G-25 column to remove the last traces of unreacted parathion and its noncovalently bound metabolites. The protein fraction from the Sephadex column was reduced in volume and subjected to SDS-polyacrylamide gel electrophoresis in the absence of either dithiothreitol or mercaptoethanol. The results are shown in Figure 5. There was considerable protein and radioactivity at the origin. This material at the origin represents an aggregate of reductase... 

Supplement 250 pi of antibody solution (about 2 mg of antibody per milliliter of Soln. A) with 1.1 pi of Soln. B and shake protected from light at RT for 1 h. Dialyze the reaction mixture three times with a 100-fold volume of Soln. A at 4 °C for 6 h each. 

Weigh 4 mg of a monochloroacetylglycyl peptide (MCA-Gly peptide, peptide carrying a MCA-glycyl residue at the N-terminus) into an Eppendorf tube and add the activated KLH or other iminothiolane-activated carrier protein. Shake vigorously at RT for 3 h. Dialyze the reaction mixture twice at RT against PBS for 1 h each. Calculate protein concentration from 235-, 260-, and 280-nm readings (cf. Protocol 1.1.7). 

Continue stirring at RT for 20 min and dialyze twice against 100 voL Soln. B each for 3 h. Transfer the dialysate into a fresh reaction tube, add quickly 10 ql Soln. C, vortex, and add immediately 5 mg antibody in Soln. D (about 10 mg/ml). 

The reaction is terminated with a small volume of aqueous, 1% hy-droquinone solution and a volume of water equal to 1/3 of the reaction solution volume is added to the product. This solution is added to 10 times its volume of 2-propanone and the polymer is recovered by filtration. The solids are redissolved in water. To remove calcium ion from the product, an amount of Na2C2C>4 equal to the moles of CaCl2 added to the reaction is placed in the solution. The CaC2C>4 precipitate is removed by filtration. The filtrate is dialyzed against distilled water for 3 to 5 days using 6 Spec-tropore dialysis tubing. The dilute aqueous solution is then freeze dried to recover the product. 

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