what is the best way to solubilize fibronectin? - (Oct/04/2006 )
Just wondering, I have lyophilized or powder..not sure at the moment....human fibronectin from BD which I would like to solubilize and put onto plates for adhesion...problem...I could find no simple technical data on how to do it or what to do it in or any other tips....hoping ya'll have some
I have coated TC slides with fibronectin before. I just put it in PBS and vortexed the holy daylights out of it, but it remained pretty 'sticky'
all right...tried to look for an old post with details and couldn't find it. this is what Sigma says:
"Fibronectin (Product Nos. F 1141*, F 0895*, F 4759, F 2006, F 0635)
Optimal conditions for attachment must be determined for each cell line and application.
Reconstitute with 1 ml sterile H2O/mg of protein. Allow to dissolve for at least 30 minutes at 37°C. A small amount of undissolved material may remain. This will not affect product performance.
Dilute fibronectin in sterile balanced salt solution and coat the culture surface with a minimal volume.
Allow to air dry for at least 45 minutes at room temperature. Excess fibronectin may be removed by aspiration, but is not necessary.
*Note: Step 1 is not necessary for the Fibronectin Solutions, Product Nos. F 1141 and F 0895.
Freezing and thawing of reconstituted fibronectin is not recommended as breakdown of protein will occur.
I never could get it to go well into solution, but apparently heating it is supposed to help?? good luck