Protein Laboratories Rehovot Ltd.
November 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 !!!
Fish gilthead seabream (Sparus aurata) IGF-I - Cat no. GHP-10 GENE ID not reported
Description: Gilthead seabream (Sparus aurata) insulin-like growth factor-I (gsIGF-I) cDNA coding the mature protein was expressed in Escherichia coli. The expressed protein contained within the inclusion-body pellet was solubilized and purified to over 98% purity, as a monomeric methionyl-gsIGF-I. Amino acid composition and N-terminal sequence confirmed the identity to be the predicted protein. For reference see Fine et al., J Endocrinol. 1997 53:139-150.

Source: E. coli. 
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Physical Appearance: White lyophilized (freeze-dried) powder 
Formulation: The protein was lyophilized from a concentrated (1mg/ml) solution with 0.02% NaHCO3.

Solubility: It is recommended to reconstitute the lyophilized gsIGF-I in sterile 0.4% NaHCO3 adjusted, not less than 100µg/ml, which can then be further diluted to other aqueous solutions.

Stability: Lyophilized gsIGF-I although stable at room temperature for several weeks, should be stored desiccated below -18 C. Upon reconstitution at > 0.1 gsIGF-I mg/ml and up to 2 2 mg/ml and filter sterilization gsIGF-I can be stored at +4C. 
Purity: Greater than 98.0% as determined by:
(a) Analysis by reducing and non-reducing SDS-PAGE gel.
(b) Gel-filtration chromatography under non denaturing conditions.
Amino Acid Sequence: The sequence of the first ten N-terminal amino acids was determined and was found to be Met-Ser-Pro-Glu-Thr-Leu-Cys-Gly-Ala-Glu.  
Dimers and Aggregates: The purified gsIGF-I consists of > 97% monomers as determined by gel-filtration chromatography.
Biological Activity: Binding assays of the 125I-gsIGF-I to gilthead seabream or carp (Cyprinus carpio) sera resulted in high specific binding, indicating the existence of one or more IGF-binding proteins. In binding experiments to crude gilthead seabream brain homogenate, using human (h) IGF-I as a ligand, the respective IC50 value of hIGF-I was about fourfold lower than that of gsIGF-I. Recombinant gsIGF-I exhibited mitogenic activity in a mouse mammary gland-derived MME-L1 cell line which was approximately 200-fold lower than that of hIGF-1. Binding experiments to intact MME-L1 cells suggests that this difference most likely results from a correspondingly lower affinity for IGF-I receptor in these cells. In contrast, the activities of gsIGF-I and hIGF-I measured by 35S uptake by gill arches from the goldfish (Carassius auratus) were identical, indicating that the recombinant gsIGF-I is biologically active. 
Endotoxin: Less than 0.1 ng/µg (IEU/µg) of Gilthead Seabream IGF-I 
Protein content: Protein quantization was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.60 as the extinction coefficient for a 0.1% (1mg/ml) solution at pH 8.0. This value is calculated by the DNAman computer analysis program of protein sequences.
Usage: This material is offered by PLR for laboratory research 
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