Hello, If you are working out conditions for an anti-HA-tag antibody, you must be working with cells transfected with an HA-tag-containing cDNA seq. of your protein of interest - therefore - are you sure your transfections have been successful to start with? In addition, as well as looking for the right anti-HA-tag antibody, it may be good to think about a) generating cells that stably express your protein of interest; b) adding more than one HA-tag sequence to your cDNA to amplify the signal; c) adding a different tag (a flag-tag) to your cDNA seq. 

Dear Mirsada,

Thanks for your suggestions. I already have checked that my protein contains the HA epitope tag by WB. My issue is related with the Immunofluorescence, the anti-HA that I have used give me plenty of noise signal.

Cheers,

Manuel

ab9110 from abcam is very good. I use it at 1:1000 in 3%BSA PBS for 2 h at RT

Hope it helps

Francisco

Thanks, Francisco.

I will try it.

Manuel

Rabbit anti-HA (invitrogen 71-5500) is really good in immunofluorescence.

Bruno

12CA5 mouse monoclonal also works well. It is available from a number of companies.

Roche, Rat anti-HA High-affinity monoclonal ( Clone 3F10). Lot 1564900.