Amplification with antibody stacking method, Binding Site kit

<< Previous Message | Next Message >>
From:Gayle Callis <>
Content-Type:text/plain; charset=us-ascii

Has anyone played with the polyMICA detection kit from Binding Site based
on stacking of secondary antibodies

Primary mouse antiWhatever!
Secondaries sequencing

Sheep antimouse
Donkey antisheep HRP
Sheep antiDonkey
Donkey antisheep HRP,

I find this very interesting, have discussed it a bit with others on the
ability of this method to amplify the detection of low level antigens other
than a tyramide technology. 
There are similar amplification methods (double cycling of secondary and
substrate or repeating the secondary/substrate steps), basically the same
here, except biotin and avidin are taken out of the loop.    

Has anyone tried this using inhouse dilutions of  secondary antibodies,
rather than the kit? 

Have a good weekend

Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

<< Previous Message | Next Message >>