Description:
Recombinant Placental Human GH-20K produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 177 amino acids and having a molecular mass of 20498 Dalton. Predicted pI=8.20. rHuGH-20K-pl is almost completely devoid of lactogenic (prolactin receptor mediated) activity characteristic to pituitary hGHs.
rHuGH-20K-pl is purified by proprietary chromatographic techniques.
Source:
Escherichia coli.
Physical Appearance:
Sterile filtered white lyophilized (freeze-dried) powder.
Formulation:
The protein was lyophilized from a concentrated (1mg/ml) solution with 0.0045 M NaHCO3 previously adjusted pH 11.
Solubility:
It is recommended to reconstitute the lyophilized rHuGH-20K-pl in 0.4% NaHCO3 or water adjusted to pH 11, not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Stability:
Lyophilized rHuGH-20K-pl although stable at room temperature for 3 weeks, should be stored desiccated below -18C. Upon reconstitution and filter sterilization rHuGH-20K-pl can be stored at 4C, pH 11 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 11.
(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-Phe-Pro-Thr-Ile.
Dimers and Aggregates:
Less than 3% as determined by silver-stained SDS-PAGE gel analysis.
Biological Activity:
PLR’r HuGH-20K is fully biologically active when compared to World Health Organization (WHO) reference standard as evidenced by in vitro bioassay. It is also capable of forming a 1:2 complex with the recombinant human growth hormone receptor extracellular domain (ECD).
Endotoxin:
Less than 0.1 ng/µg (IEU/µg) of human GH.
Protein content:
Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 1.08 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by DNAman computer analysis program of protein sequences.
Usage:
This material is offered by PLR for research, laboratory or further manufacturing purposes.
