The level of coating will vary for different species and types of cells. In general final coating concentrations from 3-10µg/cm(2) of surface area will typically supply sufficent coatings for most cell types.
Surface area of plates: (pi)(Radius squared) in cm(2)
In partical terms working dilutions of 50µg/mL to 100µg/mL are usually adequate.
Dilute poly-D-lysine in PBS, water, or 0.1M borate buffer {prepared by adding 1.24 g boric acid and 1.9 g sodium tetraborate in 400 ml water, pH 8.5} to final working concentration.
Plates or flasks are coated for 3 hours to overnight, and up to 3 days if stored at 4C. Best results are usually with just over night coatings.
Remove poly-d-lysine solution and rinse with sterile PBS or water 1-2X. Do not scrape the bottom of the coated plates or flasks.
Use plates immediately or store with PBS for up to 5 days.
Poly-D-lysine coating methods will vary slightly from laboratory to laboratory, please use the method as a general guide.
General description
Poly-D-Lysine promotes the adhesion of tissues/sections to the culture vessel. The polycationic property of the molecule allows interaction with anionic sites on the tissue/cells promoting effective attachment to the growing surface. The molecular weight of Poly-D-Lysine is >300,000 Da.