I have been using conjugated antibodies for staining cancer cells. And I try preserving the fluorescence signals using Prolong antifade with Dapi. However, after 24-48 hours, the only signal I see is DAPI. All other signals such as FITC (cytokeratin) and TRITC (CD45) are lost/ barely visible. Is this a common problem with conjugated antibodies?
In indirect immunofluorescence technique, multiple secondary antibodies will bind to the primary antibody used to bind the protein of interest. This allows us to get a better signal than just using a conjugated primary antibody. In your case, looks like the problem comes from photobleaching of FITC and TRITC ( I am assuming you were getting good fluorescence initially but lost after preservation). I think you can try alexa conjugated antibodies instead of regular FITC or TRITC. I have used alexa 488, alexa 647 and alexa 568 conjugated antibodies. I found them preserving fluorescence signal quite well.
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