Pierce™ EDC, No-Weigh™ Format

No. EDC is very unstable in aqueous environments and must be dissolved immediately before use. NHS and Sulfo-NHS are relatively stable in solution but best results are obtained when they are dissolved immediately before use. Store these compounds desiccated at 4°C.

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These compounds are used to modify a carboxyl group to form an amine-reactive ester. This is accomplished by mixing NHS with a carboxyl-containing molecule and the carbodiimide EDC (Cat. No. 22980, 22981, 77149, A35391). EDC causes a dehydration reaction between the carboxyl and the NHS hydroxyl group, giving rise to an NHS-ester-activated molecule. The activated molecule can then be reacted spontaneously with a primary amine-containing molecule. Although the carboxyl-molecules can be made to react directly with amines using EDC, the reaction is much more efficient with NHS because a stable intermediate is created. In fact, molecules that are activated as NHS esters can be dried and stored for later reaction to amine-containing targets.

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These compounds are used in conjunction with the crosslinker EDC (Cat. No. 22980, 22981, 77149, A35391) to synthesize amine-reactive labeling reagents, crosslinkers and conjugation compounds. Any compound containing a carboxylic acid (-COOH), such as a protein, or biotin or peptide, can be activated with NHS or Sulfo-NHS to form an NHS ester that will spontaneously react to form covalent amide linkages with proteins and other molecules that contain primary amines (-NH2).

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The CarboxyLink Resin is for immobilizing carboxyl-containing biomolecules after EDC activation. CarboLink Resin is hydrazide-activated and immobilizes glycoproteins. Carbolink has been replaced with glycolink immobilization kits and resins. Glycolink can also be used to immobilize steroids or other molecules containing ketones. Both resins are capable of coupling ligands via carboxylic acids using EDC (Cat. No. 22980). Both immobilization chemistries are available on UltraLink Resin as UltraLink Hydrazide (glycoproteins) and UltraLink DADPA (carboxyl) Resins.

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These resins are derivatized with diaminodipropylamine (DADPA). DADPA has a terminal primary amine that enables covalent immobilization of peptides or other carboxyl-containing (-COOH) molecules. When incubated with the resin and the carbodiimide crosslinker EDC (included in the CarboxyLink Immobilization Kit), carboxyl-containing molecules become permanently attached to the resin by stable amide bonds. The immobilized molecules can then be used in affinity purification procedures. CarboxyLink/DADPA UltraLink Coupling Resins can also be used to immobilize other kinds of molecules using alternative amine-reactive crosslinking chemistries.

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