[Histonet] Strange circles in IHC slides

[Anthony Reilly]

Hi
 
Looking at the images this appears to be the prozone effect which is caused by the primary antibody concentration being too high and has been well reported in both immunohistochemistry, immunology and serology articles.  Do a google serch for prozone phenomenon and you will find an explanation of the theory involved.    The classic feature are the circular areas which are unstained and this is reinforced in the images by the amount of background staining in the areas that are stained.  Try titrating your primary out and I am sure will eliminate the problem.
 
regards
 
 
Tony Reilly

Chief Scientist
Anatomical Pathology
Pathology Queensland
Level 1, Building 15
Princess Alexandra Hospital
Ipswich Rd, 
Woolloongabba Q 4102
Australia
Ph: 07 32402412
Fax:07 32402930
tony_reilly <@t> health.qld.gov.au


>>> "V. Neubert" <histonet.nospam <@t> vneubert.com> 3/04/2009 2:12 am >>>
Hello,

I really have some real links to real pictures of real relevance of the 
real histonet list. Really promised.

http://img12.imageshack.us/img12/8513/ts0402162049.jpg 
http://img13.imageshack.us/img13/6514/ts0402162104.jpg 

So, has ever anyone experienced sth. like this?
My conjugate control (every step except the antibody) was fine, nothing 
to be seen about DAB and no circles at all.

I used Shandon single-use coverplates, sterile buffer, fresh antibody 
aliquots. Any idea?

Thanks,

V. Neubert

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