How do you make poly-lysine coated slides?
How do you make poly-lysine coated slides?
Coating Procedure:
- Allow diluted Poly-L-Lysine solution to come to room temperature (18-26°C) before use.
- Place clean slides, a rack at a time, in diluted Poly-L-Lysine solution for 5 minutes.
- Drain slides and dry in 60°C oven for one hour or at room temperature (18-26°C) overnight.
How do you dilute poly-L-lysine?
Dilute Poly-L-Lysine Solution 1:10 with deion- ized water prior to coating slides. Use plastic con- tainers and graduated cylinders when mixing or storing solution and coating slides. Do not add fresh solution to used diluted solution.
How do you make poly lysine solution?
To Prepare Stock Solution: Dissolve 10 mg poly-L-lysine in 1 ml sterile, deionized water. This creates a 1% solution. Best to use a product with a molecular weight not less than 30,000. Optimal molecular weight should be determined by each individual lab.
How do you take Poly D lysine?
- Dissolve 5mg poly-d-lysine, (70-150kd molecular weight), in 100ml sterile water to 50ug/ml.
- Add enough poly-d-lysine solution to cover culture surface.
- Incubate 1-24 hours.
- Remove polylysine solution and rinse the plate thoroughly.
- Let the plate dry.
- Store coated plates room temperature or 4-8°C.
How do you store poly-L-lysine plates?
Storage: Store coated plates at 4-8* or room temperature. Store PDL solution at 2-8* for shorter periods of time. Store at -20* for extended storage times.
What is poly-D-lysine used for?
Poly-D-Lysine is a chemically synthesized extracellular matrix used to facilitate cell adhesion to tissue culture-treated plastic and glass surfaces.
How do you apply polylysine coating?
What is the protocol for coating surfaces with poly-D-lysine?
The protocol for coating surfaces with poly-D-lysine is the same as for poly-L-lysine. The optimal conditions must be determined for each cell line and application. In general, the following steps can be used.1. Add 50 ml of sterile tissue culture grade water to 5 mg of poly-D-lysine.2.
How do I prepare poly-D-lysine hydrobromide?
Poly-D-lysine is recommended since it is not subjected to protease released from cells. Prepare 1mg/ml stock solution by dissolving 100mg Poly-D-lysine hydrobromide(Sigma P-0899) in 100 ml water. Filter sterilize the solution then store aliquots at –20℃.
How to dissolve poly-D-lysine in PBS?
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. More
How do you prepare polylysine coating solution?
Cover culture surface with polylysine coating solution. Incubate 1 hour at room temperature in hood. Remove polylysine solution and rinse 3 time with PBS. Dissolve 10mg poly-l-lysine in 1ml water as 1% stock solution. Dilute stock solution 2 fold in PBS as 1x coating solution.
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