Edc and nhs activation

Issue 86, From the journal: RSC Advances. You have access to this article. Please wait while we load your content Something went wrong. Try again? Cited by. Download options Please wait Supplementary information PDF K. Article type Communication. Submitted 14 Jul Accepted 03 Aug First published 03 Aug Download Citation. RSC Adv. Author version available. Download author version PDF. Request permissions. Social activity. Search articles by author Qin Yan. Hong-Ning Zheng.

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.

The O-acylisourea intermediate is unstable in aqueous solutions and failure to react with an amine will cause hydrolysis of the intermediate, regeneration of the carboxyls, and the release of an N-substituted urea. Therefore, it is necessary to quench the EDC activation reaction with a thiol-containing compound like 2-mercaptoethanol. EDC couples NHS to carboxyls, which forms an NHS ester that is considerably more stable than the O-acylisourea intermediate and allows for efficient conjugation to primary amines at physiologic pH.

Topics: Cross-Linkers. This assay is suitable for the simple and rapid estimation of protein concentration. This assay is based on a single Coomassie dye based reagent. The binding of protein to the dye results in a change of color from brown to blue.

The change in color density is proportional to protein concentration. Protein estimation can be performed using as little as 0. NOTE: These are highly hygroscopic; failure to equilibrate may lead to poor cross linking.