Thank you very much for your help!!....
I read and also got in contact with Chemicon company and they also said is because I am Not using Glutaraldehyde in fixation solution. I perfused with 0.9% Saline and 4% Paraformaldehyde
I do IHC, each antibody separetly, not in combination..using same brain, buffers solutions and protocol...can I do different antibody in same buffer solution and run at the same time??..
Thank you for your caring!.



Luz Cardenas, MD
Post-doctoral Research Trainee
Department of Anatomy and Neurobiology
University of Tennessee Health Science Center
855 Monroe Avenue, Room 309/302
Memphis, TN 38163
Lab: (901) 448-6002 / 7318 
Fax: (901) 448-7193