Certificate of Analysis & Data Sheet

IGF-1 is a small protein secreted mainly but not solely by the liver and circulating in blood mostly as a complex with various IGF binding proteins in the blood. It has growth-regulating, insulin-like, and mitogenic activities and it is secreted in response to growth hormone stimulation. Recombinant rabbit IGF-I produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 71 amino acids and having a molecular mass of 7639 Dalton.

E.coli

White lyophilized powder

The protein was lyophilized from a concentrated (0.5 mg/ml) solution with 0.02% NaHCO3.

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

Lyophilized rbIGF-I although stable at room temperature for several weeks, should be stored desiccated below -18 C. Upon reconstitution at > 0.1 rbIGF-I mg/ml and up to 2 2 mg/ml and filter sterilization rbIGF-I can be stored at +4C.

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.

MTAGPETLCGAELVDALQFVCGDRGFYFNKPTGYGSSSRRAPQTGIVDECCFRSCDLRRLEMYCAPLKP

The purified rbIGF-I consists of > 95% monomers as determined by gel-filtration chromatography.

rbIGF-I is biologically active when compared to human IGF-1. The ED50, calculated by the dose -dependent proliferation of human MCF/7 cells is 5 to 25 ng/ml in the cell culture mixture dependent on culture conditions. Its activity consisits of 30-40 % compared to human IGF-1.

Endotoxin level is <0.5 EU/ug.

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.

This material is offered by PLR for laboratory research only.