Hi,

In my hands the "classic" anti-HA antibody 12CA5 worked, but only just, even with a 3xHA tag.

I had very good results with a rabbit anti-HA (sc805 from Santa Cruz) at 1:5000 dilution.

If you need a monoclonal, you could try the the newer (rat?) 3F10 clone, I haven't used it in Dicty, but at least in plants, it worked much better than 12CA5 (and I tried batches of 12CA5 from a few different companies) 

I hope this helps,

manu dubin


--- On Fri, 8/20/10, Ted Cox <[log in to unmask]> wrote:

> From: Ted Cox <[log in to unmask]>
> Subject: [DICTY - 390] HA tags
> To: [log in to unmask]
> Date: Friday, August 20, 2010, 4:28 AM
> Dear All,
> 
> We've had trouble detecting a single C-terminal HA tag in
> transformed cells, both in cell extracts, and to a lesser
> extent, with partially purified proteins.  We think it
> may be the primary antibody--there are many sources. 
> Any observations and help would be greatly appreciated.
> 
> Ted
> 
> Edward Cox
> Professor of Molecular Biology
> Edwin Grant Conklin Professor of Biology
> 333 Moffett Lab
> Phone: 609-258-3856
> Lab Phone: 609-258-3571
> Fax: 609 258 6730
> 
> [log in to unmask]
> http://www.molbio2.princeton.edu/index.php?option=content&task=view&id=206
>