What is EDC NHS chemistry?

What is EDC NHS chemistry?

EDC (also called EDAC) is 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide hydrochloride, MW 191.70. DCC is N’, N’-dicyclohexyl carbodiimide, MW 206.32. Carbodiimide conjugation, as with CDI-mediated conjugation, works by activating carboxyl groups for direct reaction with primary amines via amide bond formation.

How do you remove N hydroxy succinimide?

As Larissa commented, for future reactions you should use EDCI, whose urea-based biproduct is easily removed by sequential acid washes, or DCC, which typically precipitates out of organic reactions as an insoluble urea, thus is easily removed by filtration.

How do you store sulfo NHS?

Sulfo-NHS is water soluble and long-lived and hydrolyzes relatively slowly in water and aqueous buffer, unlike NHS. It is shipped at ambient temperature. Upon arrival, store it refrigerated at 4°C. This product is stable for 1 year at 4°C.

What does EDC and NHS do?

EDC, in conjunction with NHS allows, for 2-step coupling of two proteins without affecting the carboxyls of the second protein. First, EDC activates carboxyl groups and forms an amine reactive O-acylisourea intermediate that spontaneously reacts with primary amines to form an amide bond and an isourea by-product.

Does EDC react with water?

EDC has a high solubility in water (> then 200g/l) and is also soluble in a variety of organic solvents e.g. dichloromethane, tetrahydrofuran and dimethylformamide. Although EDC can be used for reactions in aqueous solution it is usually stored under dry conditions to prevent slow hydrolysis to the corresponding urea.

Is NHS a good leaving group?

As NHS is highly reactive at physiological pH, it is used for amine coupling reactions in bioconjugation. As shown in Fig. 8.5, the NHS ester compounds react with nucleophiles to form an acylated product with NHS as a leaving group. Carboxyl groups activated with NHS esters are highly reactive with amine nucleophiles.

What is the difference between NHS and sulfo-NHS?

Both NHS and Sulfo-NHS are soluble in aqueous and organic solvents. Activation with NHS, however, decreases water- solubility of the modified carboxylate molecule, while activation with Sulfo-NHS preserves or increases water-solubility of the modified molecule, by virtue of the charged sulfonate group.

Can you provide the shelf life for sulfo-NHS (N-hydroxysulfosuccinimide)?

Can you provide the shelf life for Sulfo-NHS (N-hydroxysulfosuccinimide)? Sulfo-NHS is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended.

Is NHS or sulfo-NHS required for carbodiimide reactions?

Although NHS or Sulfo- NHS is not required for carbodiimide reactions, their use greatly enhances coupling efficiency. Furthermore, using NHS or Sulfo-NHS makes it possible to perform a two-step reaction. Both NHS and Sulfo-NHS are soluble in aqueous and organic solvents.

What is sulfo-NHS reagent?

Thermo Scientific Pierce Sulfo-NHS is a chemical modification reagent for converting carboxyl groups to amine-reactive NHS esters for bioconjugation, crosslinking, labeling and immobilization methods. Features of Sulfo-NHS: • Efficiency of EDC-mediated coupling is increased in the presence of Sulfo-NHS.