Protein Laboratories Rehovot Ltd.
May 2019

Certificate of Analysis & Data Sheet

We are the ONLY company producing leptin antagonists and leptins of farm animals including several fish species.

Pegylated human, mouse, rat and ovine leptins and corresponding leptin antagonists are now available!!!

PLR provides a novel service of pegylation of either existing, or provided or custom-made proteins. All PLR's proteins are carrier free (cf) and can be sold in any requested amount !!!
Ovine growth hormone antagonist (G119R) - Cat no. GHP-4 GENE ID 443329
Description: Recombinant ovine GH-22K G119R produced in E. coli is a single, non-glycosylated, polypeptide chain containing 191 amino acids and having a molecular mass of 22 kDa. It is mutated in single amino acid (G119R) according to similar bovine GH mutein described by Chen et al. (1991), Molecular Endocrinology 5:1845–1852. oGH was purified by proprietary chromatographic techniques.

Source: Escherichia coli.

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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 9.
Solubility: It is recommended to reconstitute the lyophilized oGH G119R 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 oGH G119R although stable at room temperature for at least two 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-Ala.
Dimers and Aggregates: Less than 4% as determined by gel filtration chromatography
Biological Activity: PLR’s oGH G119R acts as antagonist using an in vitro bioassay in PDF-P1 3B9 cells stably transfected with rabbit GH receptors. It is capable of forming a 1:1 complex with the recombinant ovine growth hormone receptor extracellular domain (ECD)and binds to this ECD with affinity similar the the wild type oGH. 
Endotoxin: Less than 0.1 ng/µg (IEU/µg) of ovine GH G119R. 
Protein content: Protein quantitation was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.63 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by DNAman computer analysis program of protein sequences.  
Usage: For research only 
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