protein

Rat growth hormone (rGH)-

GHP-6

Recombinant rat GH-22K produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 192 amino acids and having a molecular mass of 22367 Dalton. Predicted pI=6.36. rGH is purified by proprietary chromatographic techniques.

Escherichia coli

White lyophilized (freeze-dried) powder.

The protein was lyophilized from a concentrated (1mg/ml) solution with 0.0045 M NaHCO3 previously adjusted pH 8-9.

It is recommended to reconstitute the lyophilized rGH in 0.4% NaHCO3 or water adjusted to pH 9, not less than 100µg/ml, which can then be further diluted to other aqueous solutions, preferably in a presence of a carrier protein such as BSA or similar.

Lyophilized rGH although stable at room temperature for 3 weeks, should be stored desiccated below -18C. Upon reconstitution and filter sterilization rGH can be stored at 4C, pH 9 for up to 4 weeks. For long term storage and more diluted solutions it is recommended to add a carrier protein (0.1% HSA or BSA). Please prevent freeze-thaw cycles.

Greater than 98.0% as determined by: (a) Analysis by RP-HPLC. (b) Gel filtration at non denaturing conditions using 25 nM Tris-HCl + 150 nM NaCl, pH 8. (c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Phe-Pro-Ala-Met.

Less than 5% as determined by gel filtration chromatography

PLR’s rGH is fully biologically active when compared to World Health Organization (WHO) reference standard using in vitro bioassay. It is also capable of forming a 1:2 complex with the recombinant ovine growth hormone receptor extracellular domain (ECD).

Less than 0.05 ng/µg (IEU/µg) of rat GH.

Protein quantitation was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.69 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by DNAman computer analysis program of protein sequences.

For research only