Fibronectin coating - (May/18/2010 )

Hi, I'm gonna coat my plates with fibronectin and first of all I want to know if what I've plan to do is ok:

Coat plates with 10µg/ml fibronectin in PBS1x. Incubate ON at 4ºC.

Rinse plates once with PBS 1x.

Pre-warm plates at 37ºC before seeding.

And secondly, I wonder if I can reuse fibronectin solution used for coating and if so, how to keep it.

Thanks¡¡

Hi.

What we generally do, is to add 1mL of a 10µg/mL Fibronectin solution (fibronectin diluted in PBS 1x pH7.4) to each well, and then incubate at 37C 5% CO2 for 1h. Then we rinse the plates with PBS (1x pH7.4), before adding the cells. This cuts the time you need for cell culture preparations by a lot.

Ace