I have performed few of the initial experiments for HUVEC cell migration in BD 24 well FluorBlock fibronectin coated plates. I procured the pre-screened primary HUVEC cells from Promocell. I am able to get 8-10 fold migratory response with 5%FBS after 24hrs but only around 2 fold migration with 5ng/mL VEGF when detected with Flexstation-III after Calcein staining. Tried overnight fasting of cells before the assay. Also increase in VEGF concentration didn't increase the migration. Please suggest how can I get better response?

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