Recombinant murine IGF- produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 70 amino acids and having a molecular mass of 7600 Dalton.
rmIGF- is purified by proprietary chromatographic techniques.
| Physical Appearance:
|| Sterile Filtered White lyophilized (freeze-dried) powder.
|| The protein was lyophilized after extensive dialysis against 0.2µm-filtered solution in PBS.
It is recommended to reconstitute the lyophilized rmIGF-I in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
|| Lyophilized rmIGF-I although stable at room temperature for 3 weeks, should be stored desiccated
below -18 °C. Upon reconstitution rmIGF-I should be stored at 4 °C between 2-7 days and for future use
below -18 °C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please prevent freeze-thaw cycles.
|| Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(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 Gly-Pro-Glu-Thr-Leu.
N-terminal methionine has been completely removed enzymatically.
| Dimers and aggregates:
|| Less than 1% as determined by silver-stained SDS-PAGE gel analysis.
| Biological Activity:
|| ProSpec's rmIGF- is fully biologically active when compared to standards.
The ED50, calculated by the dose-dependant proliferation of murine BALB\C 3T3 cells (measured by 3H-thymidine uptake) isless then1.0 ng/ml, corresponding to a specific activity of 1x106 U/mg.
|| Less than 0.1 ng/µg (IEU/µg) of rmIGF-I .
| Protein content:
|| Protein quantitation was carried out by two independent methods:
UV spectroscopy at 280 nm using the absorbency value of 0.47 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). Analysis by RP-HPLC, using a calibrated solution of IGF- as a Reference Standard.
This material is for research, laboratory or further manufacturing purposes