Thrombin Sepharose beads are designed for efficient cleavage of recombinant fusion proteins, containing a thrombin cleavage site (e.g. Leu-Val-Pro-Arg-ll-Gly-Ser; ll: cleavage site), circumventing the need for chromatographic techniques to remove thrombin after cleavage reaction is completed. BioVision’s Thrombin Sepharose is prepared by covalent coupling of bovine thrombin to activated 6% cross-linked Sepharose beads. 15 µl of the slurry is sufficient to cleave >90% of 1 mg of BioVision’s 6xHis-tagged fusion protein in 50 mM Tris buffer, 0.1 M NaCl, pH 8.0 within 4 h at room temperature. These beads can be regenerated for repeat use. The reusability depends on multiple criteria including the usage and handling of the product.