Description:
Recombinant ovine GH-22K produced in E. coli is a single, non-glycosylated, polypeptide chain containing 191 amino acids and having a molecular mass of 21980 Dalton. Predicted pI=7.81. oGH is purified by proprietary chromatographic techniques. It differs from pituitary oGH by two amino acids G9R/G63S and possesses higher biological activity as compared to pituitary oGH. For ref see Reicher et al. (submitted)
Source:
E. coli
Physical Appearance:
Lyophylized white powder
Formulation:
The protein was lyophilized from a concentrated (1mg/ml) solution with 0.0045 M NaHCO3 previously adjusted pH 9.
Solubility:
It is recommended to reconstitute the lyophilized oGH 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.
Stability:
Lyophilized oplGH although stable at room temperature for 3 weeks, should be stored desiccated below -18C. Upon reconstitution and filter sterilization oGH 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.
Purity:
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.
Amino Acid Sequence:
The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Thr-Phe-Pro-AlaLess than 4% as determined by gel filtration chromatography
Dimers and Aggregates:
Less than 4% as determined by gel filtration chromatography
Biological Activity:
PLR’s oplGH is fully biologically active when compared to World Health Organization (WHO) reference standard using in vitro bioassay in PDF-P1 3B9 cells stably transfected with rabbit GH receptors. It is also capable of forming a 1:2 complex with the recombinant ovine growth hormone receptor extracellular domain (ECD).
Endotoxin:
Less than 0.1 ng/µg (IEU/µg) of oplGH (22 K).
Protein content:
GH.Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 0.684 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 calibrated solution of GH-22K-pl as a Reference Standard.
Usage:
For research only
