多聚赖氨酸包被方法

Poly-lysine-coated tissue culture surfaces

1. To coat coverslips:

Prepare a stock solution by dissolving 25 mg/ml polylysine in ml water (both poly-L-lysine and poly-D-lysine are used to coat tissue culture surfaces; check specific protocol for choice of is omer) and filter sterilize through a μm filter.

Store in 100-μl aliquots at -20°C.

When ready to use, dilute one aliquot in 40 ml water to prepare 13 μg/ml working solution.

Sterilize coverslips by autoclaving prior to coating.

Dip coverslips in the working solution, then incubate 15 min to several hours in a humidified 37°C, 5% CO2 incubator.

Allow surface to dry.

2. To coat culture dishes: 多聚赖氨酸包被培养皿/板

Prepare a stock solution by dissolving 100 mg poly-lysine in 100 ml water (both poly-L-lysine and poly-D-lysine are used to coat tissue culture surfaces; check specific protocol for choice of isomer) and filter sterilize through a μm fil ter.

配制1mg/ml储存液

Store in 5-ml aliquots at -20°C.

分装保存在-20度

When ready to use, dilute 1 part stock solution with 9 parts water to prepare 100 μg/ml working solution.

使用前按1：10稀释成100μg/ml工作浓度。用5-10ml一次性无菌注射器和μm 滤器过滤除菌，备用。

Fill tissue culture dishes or slide wells with the working solution and incubate 1 hr in a humidified 37°C, 5% CO2 incubator, then remove solution by vacuum aspiration and allow surface to dry.

加入培养皿或培养板中，使能完全覆盖皿底即可，37度培养箱放置1小时或置超净台内室温过夜。然后吸取液体，使表面晾干。

Store coated tissue culture ware up to 3 months at 4°C.

可以立即使用也可保存在4度3个月备用。

Use diluted solutions only once, but unused diluted aliquots can be stored up to 3 months at 4°C.

加入细胞前用1XPBS 洗三遍，然后加入细胞和培养液。