Hi Everyone.
I am looking to detect the subcellular localization of a viral protein in neurons by immunofluorescence with a HA antibody. I have always made protein fusions with 3 tandem copies of HA tag in the past (3XHA). This time I have only room for one HA copy. Would that be enough for a good commercial HA antibody to work with decent specificity and affinity?
Thank you all!
Yes, we almost always use only one HA tag and visualize it by IF just fine. We use Abcam9110 anti-HA antibody.
Is there a particular reason you need to use the HA tag? I have personally found this to be a less than reliable tag. I would recommend a 3X Flag tag, 5X Myc tag or a V5 tag over HA any day. In my experience, I believe the HA tag was modified as I could see overexpressed HA-tagged protein using an antibody to that protein, but not using an antibody to the HA tag. Very frustrating if you ask me. Best of luck!
It could be HA or other tags. I just have used HA before and has worked for me. I also need a small tag, nothing larger than ~30bp.
Great feedback, Michael.
Thanks a lot!
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