The Profinity eXact purification resin consists of a highly engineered subtilisin protease conjugated to an agarose-based matrix. Resulting purity of the eluted protein is typically higher than that for other affinity-tag systems due to the specific recognition (KD < 100 pM) that subtilisin has for its prodomain sequence (Profinity eXact tag). Once the Profinity eXact fusion protein has been captured by the Profinity eXact purification resin — and the cleavage reaction has been triggered with a sodium fluoride-containing buffer — the affinity tag is precisely cleaved at the C-terminus of the cleavage recognition sequence (EEDKLFKAL). The result is a quickly purified protein product that lacks any amino acid residues derived from the Profinity eXact tag.
Benefits of Profinity eXact resin:
- Purification and processing of fusion-tagged proteins in a single-step
- On-column cleavage in as little as 30 min
- No protease addition required
- Precise cleavage at N-terminus to generate native protein sequence
The following convenient packaging configurations allow easy evaluation of Profinity eXact technology:
- Profinity eXact resin — 10 ml resin supplied in 100 mM sodium phosphate, pH 7.2 containing 0.02% sodium azide
- Profinity eXact mini spin columns — 10-pack of single-use columns that contain 0.1 ml settled resin in addition to all required collection tubes for binding, washing, and elution steps. Fit all benchtop centrifuges
- Profinity eXact mini spin starter kit — comprehensive 10-pack kit that ensures purification success upon initial trial. Includes 10 prepacked spin columns, collection tubes, lysis reagent, buffers for purification, and lyophilized MBP control protein lysate
- Bio-Scale Mini Profinity eXact cartridges — 1 ml and 5 ml cartridges that can be used with BioLogic chromatography systems, the Profinia protein purification system, a peristaltic pump, or with appropriate column adaptors with any other chromatography system
- Profinity eXact expression and purification starter kit — includes two kits: the Profinity eXact cloning and expression starter kit for the cloning and expression of a target gene using pPAL vectors and competent cells, and the Profinity eXact mini spin purification starter kit for single-step purification and on-column cleavage of the tagged protein
