New Profinity Exact Cloning and Expression Kits for the Purification and Tag-Removal of Fusion Tagged Proteins
Product News Jul 22, 2008
Bio-Rad Laboratories, Inc. has announced the availability of bulletin (number 5646A) entitled, “An Unrivaled, Novel Tag System for the Expression of Fusion-Tagged Proteins,” which describes the features and applications of new cloning and expression kits for the Bio-Rad Profinity eXact™ fusion-tag system.
Profinity eXact cloning and expression kits are part of the Profinity eXact fusion-tag system designed to address bottlenecks in procedures for purification and tag-removal of fusion-tagged proteins.
The bulletin describes how the Profinity eXact pPAL7 expression vectors facilitate the expression of a unique N-terminal tag sequence encoding a built-in cleavage recognition site, occurring at the C-terminus of the tag.
The N-terminal Profinity eXact affinity tag is the prodomain of the Subtilisin protease. Both tag and protease have been extensively modified for optimal purification results and exhibit a high degree of affinity (KD< 100pm).
The Subtilisin protease has been cleverly immobilized onto the purification resin for binding of Profinity eXact fusion-tagged proteins. Cleavage by Subtilisin is triggered by fluoride buffers, occurs on-column and directly after the cleavage recognition site, and leaves a native protein behind.
With cleavage incubation times as short as 30 minutes, the Profinity eXact system can provide an alternative to purify and process fusion proteins without the addition of protease.