Hepatocyte growth factor (HGF), also known as Hepatopoietin A, is mitogenic for a variety of cell types, including endothelial and epithelial cells, melanocytes, and keratinocytes. It is identical to scatter factor, a fibroblast-derived soluble factor that promotes the dissociation of epithelial and vascular endothelial cell colonies in monolayer cultures by stimulating cell migration. HGF is secreted as an inactive single chain precursor and is converted to the heterodimeric active form by HGF activator.
Figure 1: HGF stimulates the 3H-thymidine incorporation by 4MBr cells in a dose-dependent manner. The ED50 for this effect is typically 20 - 40 ng/mL.
Figure 2:Typical data for the anti-HGF antibody is shown in Figure 2. To measure the ability of the antibody to neutralize the bioactivity of rhHGF on 4MBr cells, rhHGF was incubated with various concentrations of antibody for 1 hour at 37° C. Following this preincubation period, 4MBr cells were added to a microplate. The assay mixture in a total volume of 100 μL, containing antibody at the concentrations indicated, rhHGF at 100 ng/mL and cells at 1 x 105 cells/mL, was incubated for 48 hours at 37° C in a 5% CO2 humidified incubator and pulsed with 3H-thymidine for the final 24 hours. The cells were then detached and harvested onto glass fiber filters and the 3H-thymidine incorporated into DNA was determined. The ND50 of the antibody is approximately 0.2 - 0.6 μg/mL.
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