[Histonet] Immunofluorescence double labeling with two rabbitantibodies

Anatoli Gleiberman AGleiberman <@t> cbiolabs.com
Tue Oct 12 14:22:41 CDT 2010

Jackson Immunoresearch provides protocol and reagents that are useful
for solving your problem: 
I have used this approach to localize at the same sample simultaneously
two different isoforms of a transcription factor with two different
mouse monoclonals. Worked for me.

Anatoli Gleiberman, PhD
Director of Histopathology
Cleveland Biolabs, Inc
73 High Street
Buffalo, NY 14203
phone:716-849-6810 ext.354
e-mail: AGleiberman <@t> cbiolabs.com

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Aline
Sent: Tuesday, October 12, 2010 1:29 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Immunofluorescence double labeling with two

Hello all,
I using formalin fixed paraffin embedded specimens from fetuses from  
sheep to examine which cells are virally infected within the CNS.
I am using IBA-1 to identify microglial cells. The IBA-1 antibody is a  
rabbit antibody and it works really well in my specimens.
At the same time I am also using an antibody against the virus that I  
am studying that is also a rabbit Ab.
For these samples I am using double labeling to determine if cells are  
infected or not.
Both antibodies work, but I can use them together otherwise there is x- 
Does anyone has any suggestions of how to label tissues with two  
different antibodies that come from the same species.
On immunoportal someone suggested to use 4%PFA to fix the tissues in  
between the first (primary and secondary Ab) and the second (primary  
and secondary Ab), however it did not work. PFA did not block the  
previously bound Abs.
The other option that I would have would be to use IBA-1 from goat. My  
problem with this antibody coming from goat is that I can have x- 
reaction and moderate background since goat and sheep are very closely  
Please, if someone has any suggestions I would immensely appreciate!
Thank you very much!
I hope you have a wonderful day!

Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu


This communication may contain privileged information.  It is intended solely for the use of the addressee.  If you are not the intended recipient, you are strictly prohibited from disclosing, copying, distributing or using any of this information.  If you received this communication in error, please contact the sender immediately and destroy the material in its entirety, whether electronic or hard copy.  This communication may contain nonpublic information about individuals and businesses subject to the restrictions of the Gramm-Leach-Bliley Act.  You may not directly or indirectly reuse or redisclose such information for any purpose other than to provide the services for which you are receiving the information.

More information about the Histonet mailing list