Specifications and Use of R&D HGF
Source of R&D HGF
• A DNA sequence encoding the 728 amino acid residue variant of the human pre-pro-HGF was expressed in Sf 21 insect cells using a baculovirus expression system.
R&D Human HGF Molecular Mass
• Recombinant human HGF, composed of a mixture of the single chain and the disulfide-linked heterodimeric protein, migrates as an approximately 60 - 70 kDa protein in SDS-PAGE under non-reducing conditions.
R&D HGF Purity
• > 95%, as determined by SDS-PAGE and visualized by silver stain.
• < 1.0 EU per 1 μg of the cytokine as determined by the LAL method.
Activity of R&D HGF
• The biological activity of recombinant human HGF was measured by its ability to stimulate 3H-thymidine incorporation in the HGF responsive monkey epithelial cell line, 4MBr-5 (Rubin, J.S. et al., 1991, Proc. Natl. Acad. Sci. 88:415 - 419).
• The ED50 for this effect is typically 20.0 - 40.0 ng/mL.
Formulation of R&D HGF
• Lyophilized from a 0.2 μm filtered solution in 20 mM sodium phosphate buffer plus 0.5 M sodium chloride (pH 7.0) containing 50 μg of bovine serum albumin per 1 μg of cytokine.
Reconstitution of R&D HGF
• It is recommended that sterile phosphate-buffered saline containing at least 0.1% human serum albumin or bovine serum albumin be added to the vial to prepare a stock solution of no less than 10 μg/mL of the cytokine.
Storage of R&D HGF
• Lyophilized samples are stable for up to twelve months at -20° C to -70° C.
• Upon reconstitution, this cytokine can be stored under sterile conditions at 2° - 8° C for one month or at -20° C to -70° C in a manual defrost freezer for three months without detectable loss of activity.
• Avoid repeated freeze-thaw cycles.