Status:
Expert ReviewerMember since: 2012
Organization: UNESP/IBILCE
Application Area: Protein Purification
"Benzamidine Sepharose 4 Fast Flow (high sub) resin used in affinity chromatography. This resin has covalently attached benzamidine which is an inhibitor for serine proteinases. The mixture of proteins that contain serine proteinases are passed through this resin at pH 7.5-8.5. The bound proteins are eluted with buffer pH 3.0. This is an old method which has some drawbacks as some proteins like Phospholipases may also bind to this resin. In a new method that we have developed after applying the sample in buffer containing 100mM salt, the non-specific proteins are eluted with the same buffer with an additional 500mM salt. Lastly the serine proteinases are eluted by decreasing the pH to 3.0. This method has improved the affinity based purification procedure. I recommend this method for purification of serine proteinases by Benzamidine Sepharose 4 Fast Flow (high sub). The resin can be stored in 20% ethanol after use."
Benzamidine Sepharose 4 Fast Flow (high sub) is an excellent tool for removal and/or purification of trypsin and trypsin-like serine proteases, as well as, zymogens including urokinase and prekallikrein. Removal of serine proteases is easily done directly from serum, monoclonal cell supernatants and bacterial lysates.
Benzamidine Sepharose 4 Fast Flow Features: