Fibronectin coating of glass bottom dishes and glass bottom plates

Materials

Fibronectin solution

Human Fibronectin from BD Biosciences(cat# 354008) is diluted to 10-50ug/ml with Ca++ and Mg++ free PBS. The concentration should be proportional to coating concentration, i.e, make 10ug/ml solution for 1ug/cm2 coating, 50ug/ml for 5ug/cm2 coating, etc.

Make appropriate aliquots and store at -20℃.

Do not store in frost-free freezer, avoid multiple freeze thaws.

Procedure

  • Add appropriate amount of fibronectin solution to culture surface. If cells will be cultured only on the glass surface, then only the glass surface needs to be covered with the fibronectin solution.
  • Incubate for 1 hour at room temperature.
  • Carefully aspirate the remaining solution.
  • Rinse with media or PBS.
  • Fibronectin coated glass bottom dish or glass bottom plate should be used immediately.

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