Plasmin Sepharose Beads, BioVision

Supplier: BioVision
7926-5 7926-25 7926-1
10835-556EA 180.96 USD
10835-556 10835-558 10835-380
Plasmin Sepharose Beads, BioVision
Protein Purification Protein Purification Kits
Plasmin sepharose beads for efficient and convenient cleavage of recombinant fusion proteins containing plasmin-specific cleavage site.

  • Efficient and convenient cleavage of recombinant fusion proteins
  • Reactions can be scaled up to cleave the entire amount of the target protein

The formulation is Plasmin on 6% cross linked Sepharose beads, provided as 50% slurry in pure glycerol with a Ligand Density of 0.2 mg/ml of the resin.

Protocol for best use: In order to find the optimum cleavage conditions for a target protein, it is recommended to run preliminary cleavage reactions at a small scale. Successful cleavage with plasmin is dependent upon proper folding of the target protein that enables access of the plasmin recognition sequence by the enzyme. Once optimum cleavage conditions are obtained, the reaction can be scaled up to cleave the entire amount of the target protein.

The target protein should be purified to homogeneity and dialyzed against 50 mM Tris buffer, 0.1 M NaCl, pH 7.4 before setting up the cleavage reaction. First, resuspend the beads by gentle swirling. Do not vortex. Then, Aliquot 50 µl of the suspended slurry and add to 1 mg of the target protein in an Eppendorf tube. The recommended concentration of target protein is 1 mg/ml. Mix gently by inverting the tube (do not vortex) and shake on a rotary shaker at 37°C At regular time intervals spin down the tube to aliquot a test sample and freeze it immediately. At the end of the reaction, analyze the samples by SDS-PAGE. To avoid excessive cleavage, do not incubate for longer periods.

Recovery of the cleaved target protein process includes spinning down the reaction mixture for 2-3 min at 1500 x g after the fusion protein is completely cleaved. Then, remove the supernatant and wash the beads with 0.2 ml of 50 mM Tris buffer, pH 7.4. Repeat these steps to maximize the recovery of the target protein and its cleaved fragment. Further chromatography may be necessary to remove the cleaved fragments from the target protein.
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