Growth and Differentiation Factor 5 (GDF5), human recombinant (rHuGDF5)

Recombinant human GDF-5 produced in E. coli is a homodimer comprised of a non-glycosylated polypeptide chain containing 2 x 117 amino acids and having a total molecular mass of 26.8kD. To enable bacterial expression of rhGDF-5 we removed the N-terminal sequence Ala-Pro-Leu-Thr and added the Lys. GDF5 is purified by proprietary chromatographic techniques. , , Amino Acid Sequence:, The sequence of the first five N-terminal amino acids was determined and was found to be Met-Lys-Arg-Gln-Gly. Biological Activity: Human GDF-5 is fully biologically active when compared to standard. Induction of alkaline phosphatase acrtivity in ATDC5 cells: EC50 = 40Nm. Endotoxin: Less than 0.1ng/ug (IEU/ug) of GDF-5. Protein Content: Protein quantitation was carried out by two independent methods: , 1. UV spectroscopy at 280 nm using the absorbency value of 1.15 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics). , 2. Analysis by RP-HPLC, using a standard solution of GDF-5 as a Reference Standard. Storage and Stability: Lyophilized powder is stable for at least 12 months at -20°C Reconstituted product must be stored in aliquots at -20°C. For long-term storage, it is recommended to add a carrier protein (0.1% HSA or BSA). For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.