Poly-d-lysine promotes the adhesion and good growth of mammalian cells to the glass/plastic dishes. Here we describe a procedure to prepare a 1 mg/ml stock solution of Poly-d-lysine. The easiest way to prepare the solution is to buy a 5 mg poly-d-lysine (pack-size) and dissolve all of them in autoclaved sterile miliQ water without weighing it out. This stock solution can be used for coating glass coverslips to allow cells to grow adherently.
Reagents and solutions
♦ Poly-D-Lysine hydrobromide (5 mg pack) (e.g., Sigma-Aldrich #p6407)
♦ MiliQ water (autoclaved sterile)
Equipment and disposables
♦ Syringe/pipette and pipette boy
♦ Laminar flow hood
1 mg/ml Poly-d-lysine hydrobromide solution in water
Preparation of 1 mg/ml Poly-D-Lysine solution
Use appropriate personal protective equipment (lab coat, gloves, goggles, etc) for your safety and follow your institute’s guidelines.
Prior to start:
Make the laminar floor hood ready for work (switch on UV for 20 min, and then start the airflow)
Step 1: Clean all the reagents (5 mg bottle of Poly-d-lysine hydrobromide, 5 ml syringe/pipette boy, autoclaved MilliQ water bottle) with 70% ethanol and bring them inside the laminar flow hood.
Step 2: Transfer 5 ml of MilliQ water from the water bottle to the Poly-d-Lysine bottle. Mix all the content gently until all poly-d-lysine dissolves completely.
Generally, a 5 mg poly-d-lysine bottle contains a rubber lid that is sealed with an aluminum cover. You can remove some parts of the aluminum cover that can expose the rubber lid. Now fill a syringe with 5 ml MilliQ water and insert the needle in the rubber lid and inject the water into the bottle. Content can be mixed by gently swirling and inverting the bottle.
Store the poly-d-lysine solution at 4°C.