Figure 2: LOX will exchange the NH2 group with O, which creates a reactive entity for crosslinking to other tertiary amines. In the assays of LOX, H2O2 is used to oxidize reporter molecules with horse radish peroxidase as a catalyst.
Storage and Reconstitution
Briefly centrifuge to collect the product at the bottom of the tube. Reconstituting a 5 mg tube of CS-LXE01 with 100 ml of nanopure will generate a 50 µg/ml stock of LOX in the following buffer: 4M urea in PBS pH 7.4. The protein can be frozen in liquid nitrogen and stored at –70°C. It is not recommended to store at 4°C in liquid form. The lyophilized protein is stable at 4°C desiccated (<10% humidity) for 6 months.
Purity
Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 4-20% gradient polyacrylamide gel. LOX was determined to be >80% pure (see Figure 3 Below).