Polylysine-coated tissue culture surfaces


Polylysine solutions are typically used to coat glassware and plasticware  for the attachment and growth of cells.


  1. Prepare a stock solution by dissolving 100 mg polylysine in 100 ml water (poly-L-lysine or poly-D-lysine can be used; check specific protocol for choice of isomer)and filter sterilize through a 0.22-micron  filter.
  2. Store in 5-ml aliquots at -20°C.

Coating culture dishes, multiwell plates, or chamber slides

  1. When ready to use, dilute 1 part stock solution with 19 parts water to prepare a 50 ?g/ml working solution.
  2. Fill tissue culture dishes, multiwell plates, or slide wells with the working solution and incubate 1 hr in a 37°C incubator, then remove solution by vacuum aspiration and allow surface to dry.
  3. To coat coverslips: Sterilize coverslips by autoclaving or by incubating them in 95% ethanol and drying before coating. Place coverslips in a single layer in a petri dish containing working solution and incubate 1 hr at 37°C. Remove coverslips using sterile forceps and allow surface to dry.
  4. Store coated tissue culture ware up to 3 months at 4°C. Use diluted solutions only once.

Additional information

When coating with poly-l-lysine, the efficiency of coating is relatively low when prepared with distilled water due to the acidic pH of the solution that can range between a pH of 3.3-6.6. One recommendation [1] to improve coating is to use an 8.5  pH borate buffer to dissolve the lysine, with thorough rinsing after coating. In addition to the basic techniques shown here, there are a variety of approaches recommended by vendors as shown below.

  1. Dissolve 10mg poly-l-lysine in 1ml water as 1% stock solution.
  2. Dilute stock solution 2 fold in PBS as 1x coating solution.
  3. Add enough coating solution to cover culture surface completely and incubate for 5 minutes at room temperature.
  4. Remove coating solution and rinse 2 times with PBS or serum- free culture medium.
  5. Use immediately or let it dry for future use.
  1. Dissolve poly-d-lysine, (135kd molecular weight), in sterile water to 50ug/ml.
  2. Add 0.15ml/cm² of solution culture surface.
  3. Incubate 1-20 hours.
  4. Rinse 1 time and let dry.
  1. Dissolve 5mg poly-d-lysine, (70-150kd molecular weight), in 100ml sterile water to 50ug/ml.
  2. Add enough poly-d-lysine solution to cover culture surface.
  3. Incubate 1-24 hours.
  4. Remove polylysine solution and rinse the plate thoroughly.
  5. Let the plate dry.
  6. Store coated plates room temperature or 4-8°C
  1. Dissolve 100mg poly-d-lysine, (70-150kd molecular weight), in 100ml sterile water to 1mg/ml stock solution.
  2. Dilute stock to 0.1mg/ml with sterile water.
  3. Cover culture surface with polylysine coating solution.
  4. Incubate 1 hour at room temperature in hood.
  5. Remove polylysine solution and rinse 3 time with PBS.
  1. Add 50ml of sterile water to 5mg of poly lysine.
  2. Coat cell culture surface with 1ml per 25cm² culture surface
  3. Incubate 5 minutes.
  4. Remove poly lysine solution and rinse the plate thoroughly with sterile water and let it dry at least 2 hours.


[1] Should I coat coverslips in dd-water? http://www.neuvitro.com/tips-and-calculations-polylysine-coating.htm

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