Carbodiimides l-ethyl-3-

All salts and buffer components, as well as poly-D,L-lysine (PL), CBZ-protected PL, diethylene triamine penta-acetic acid (DTPA) and its anhydrides, water-soluble carbodiimide (l-ethyl-3-(dimethylaminopropyl)-carbodiimide, EDC), (SPDP), SMCC, dithiothreithol, succinic anhydride, trinitrobenzene sulfonic acid, triethylamine, bovine serum albumin (BSA), and IV-hydroxy sulfo-succinimide (HSSI) are available from Sigma (St. Louis, MO). 

Dime thy laminopropy I )-3-ethylcarbodiimide hydrochloride [l-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide hydrochloride] see entry on p. 212 in Chapter 4. 

Molecules of interest that contain free amino groups can be coupled in aqueous solution to /S-poIy(L-malate) as amides using carbodiimides such as the water-soluble l-ethyl-3(3-dimethyIaminopropyl)carbodiimide hydrochloride (EDC) [2,12,20,21]. By this method, the molecules are attached randomly. A selective amide bond formation at the carboxylate terminus can be achieved... 

The treatment of isocyanates with 3-methyl-l-ethyl-3-phosphoIene-l-oxide (59) is a useful method for the synthesis of carbodiimides in good yields. The mechanism does not simply involve the addition of one molecule of isocyanate to another, since the kinetics are first order in isocyanate and first order in catalyst. The following mechanism has been proposed (the catalyst is here represented as... 

Purify conjugate by gel filtration, dialysis or ethanol precipitation EDC = l-ethyl-3-(3-dimethylaminopropyl)carbodiimide HQ. 

Reactions of 2,3-dioxo-l,2,3,5,6,7-hexahydropyrido[l,2,3-carboxylic acids and the homologous acetic and propionic acids, prepared by basic hydrolysis of the corresponding ester, with amines, 28% NH4OH, and hydroxylamine derivatives in the presence of l-ethyl-3-[3-(dimethylamino)propyl]carbodiimide and hydroxybenztria-zole <1995BML1527>, 1995BML1533>, and in the presence of NEt3 and A, A -bis(2-oxo-3-oxazolidinyl)phosphinic... 

COOH groups of the PE (PAA or PGA) and the -NH2 moieties of the pre-loaded protein, using l-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) as a catalyst [99]. Negligible enzyme desorption (<0.1 %) is observed from cross-linked lysozyme-loaded MS spheres after exposing the samples to an aqueous solution for 48 h, while about 25 % of the immobilized lysozyme is desorbed under the same conditions when the lysozyme is not cross-linked. 

The reactions involved in an EDC-mediated conjugation are discussed in Chapter 3, Section 1.1 (Note EDC is l-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride MW 191.7 and is sometimes referred to as EDAC). The carbodiimide first reacts with available carboxylic groups on either the carrier or hapten to form a highly reactive o-acylisourea intermediate. The activated carboxylic group then can react with a primary amine to form an amide bond, with release of the EDC mediator as a soluble isourea derivative. The reaction is quite efficient with no more than 2 hours required for it to go to completion and form a conjugated immunogen. 

Weigh out 1.25 mg of the carbodiimide EDC (l-ethyl-3-(3-dimethylaminopropyl)carbodii mide hydrochloride Thermo Fisher) into a microfuge tube. 

Figure 8 Chemiluminescent (A and B) and bioluminescent (C) detections for immobilized hybridizations of PCR product. Dg, digoxigenin Bt, biotin Ad, avidin. Procedure A [30] Biotin moiety is incorporated into PCR products during the amplification reaction, using one 5 -biotinylated primer. The product is hybridized with a Dg-labeled probe and is immobilized on streptavidin-coated magnetic beads. This capture reaction is carried out for 30 min at 37°C. A permanent magnet is used to sediment the beads during washing to remove unbound DNA. By incubation with the washed beads for 45 min at 37°C, anti-Dg antibody conjugated to HRP enzyme is bound to the Dg-labeled probe, and luminol reaction is performed for CL detection. Procedure B [31] Wells of apolystyrene microtiter plate are activated with l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide, and then coated with a labeled cDNA probe complementary to an internal region of the target DNA.

This procedure for the preparation of l-ethyl-3-(3-dimethyl-amino)propylcarbodiimide and its salts is a modification of one that has been published.4 Unsymmetrical carbodiimides have also been prepared by desulfurization of the corresponding thioureas with mercuric oxide3 or by dehydration of the corresponding ureas with -toluenesulfonyl chloride in pyridine.4 Unsymmetrical 1,3-disubstituted ureas are best prepared by the reaction... 

CaM was purified from porcine brain. The purity of proteins was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing. CaM and PDE were cross-linked with l-ethyl-3(3-dimethylamino-propyl) carbodiimide (EDC) or N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) in a buffer solution of 0.1 M HEPES (pH 7.1) in the presence of 1 mM CaCl2. After a buffer solution containing 2 mM EGTA added into the reaction solution, the CaM-PDE hybrid was separated from other ingredients by gel chromatography on a Sepharose CL-6B solumn. 

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