[Histonet] Need suggestions for secondary antibody

[Gayle Callis]

Gareth, 

Your brain is fine!  We have excellent luck with goat antiRat F(ab')2,
biotin conjugate, adsorbed to mouse from Biosource/TAGO.  You can also buy
Donkey antiRat F(ab')2 biotinylated, adsorbed to mouse and other species
from Jackson Immunoreasearch. We avoid rabbit (sticky bunny!) secondaries,
and always use F(ab')2 frag of IgG rather than whole IgG secondaries to
avoid fc receptor problems/background staining.  

These secondaries run around 0.5 mg/ml, diluted to 1:250 - 

I presume you are using a Rat antimouse monoclonal?  If you have a hamster
antimouse monoclonal, then BD Pharmingens hamster cocktail of two
monoclonals is excellent OR buy a Donkey antiGolden Syrian F(ab')2
biotinylated adsorbed to mouse or Dk antiArmenian Hamster F(ab')2
biotinylated adsorbed to mouse from Jackson Immunoresearch.  You will get
crisp results with these host anti specific hamster species/Pharm hamster
cocktail secondaries compared to a goat antihamster secondary.  We
experienced very weak, diffuse results with the latter - a horror!

Good luck 

 

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)