I am using a microelectrode and breast cancer cell line (MDA-MB-231 and MCF-7). Could you share your protocol in coating the electrode surface to enhance cell adhesion?
Which concentration of Fibronectin is best one?
Is it neccessary to do SAM before coating Fibronectin? Is the CH3 or COOH group better for binding of Fibronectin?
Using a SAM will allow you to attach the fibronectin(FN) in a chosen orientation. FN has a dipole charge that results in different parts of the protein being exposed on the cell attachment side based on the surface charge of the substrate. FN also has many hydrophilic and hydrophobic domains that allow it to attach to either type of surface but with each affecting to orientation and organization of the surface differently. Using a COOH termination on the SAM will result in the a5b1 binding site being "hidden" from the cell adhesion surface. Using an NH2 termination will make that binding site more available on the cell adhesion surface. CH3 terminations are very hydrophobic while OH terminations are very hydrophilic. You don't need to use SAMs to get FN adhesion on Au, it is hydrophilic and will have approximately the same surface characteristics as an OH terminated SAM.
I am not familiar with your cell lines so I cannot advise you on which orientation of the protein to choose but my first guess would be that you want the a5b1 binding site exposed. 25ug/mL is a good starting concentration for FN and you should not need more than 100ug/mL. Higher concentrations will result in less ordered surfaces.
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