I am interested in fluorescently imunolablelling more than one protein within the focal adhesion complex (vinculin, talin, FAK, or paxillin) and was curious if anyone has tried labeling two adapter proteins on one sample? Any evident (steric) hindrances I should watch out for?
I have used antibodies to all of these proteins in different combinations and I have never found any issues of steric hindrance. Since focal adhesion have thousands (if not more) of each of these proteins it is likley not an issue for regular fluorescent imaging. For super resolution imaging I am not sure and I would suggest looking into several papers from the Claire Waterman laboratory whom have look at these structures extensively.
Thank you for taking time out to answer my question Andrew.
Yes, it is for regular confocal fluorsecent imaging.
I will go ahead and use paxillin and vinculin together on one sample then!
Will also take a look at Claire Waterman's papers to understand this more extensively.
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