Cant seem to detect HA-tag using Immunofluorescence (View forum version)



KhaleesiDany

Posted 27 October 2015 - 12:39 AM

Good day people

 

I have been trying to show that my protein is being expressed via Immunofluorescence staining of the HA- epitope in my protein sequence. I have two different proteins, i am able to detect the one but not the other, however my knockdown data does show that both my preoteins are being expressed because i do see its activity being done. What are possible reasons for HA-tag not being detected, can it be masked by something or being cleaved or not being made. im not sure,  any suggestions  please, thanks


Rosewater

Posted 27 October 2015 - 04:32 PM

I would see if it is detectible by western blot.  If you don't see the correct size band, It could be cleaved or not expressed.  If you do see the band by western, you might assume that the HA-tag is not available in IF assay, or HA Ab does not work well for IF.