Re: [Histonet] RE: Envision-AP

From:rschoon

Ian,

I've been using the Dako Envision AP kit for years but (after much trial 
and error), I have use the New Fuchsin Kit from BioGenex as my label.  
Works great when used properly and it is permanent. Avoid xylene after 
dehydration, instead usa a non-aliphatic clearing agent (I use Richard 
Allan's Clear Rite 3).  Coverslip with your choice of media.  The 
results are a nice bright red which doesn't seam to fade at least not in 
the last 7-8 years that I've used it.

Ive not used the "PowerVision that Chris mentioned and so can't comment 
on it.  I would like some information on it (better yet a trial).

Robeert Schoonhoven
UNC-CH


C.M. van der Loos wrote:

>Ian,
>I was thinking the same way as Gayle did. But apparently this wasn't the problem. I wondered what AP chromogen you are using? I have very good results using the newest Dako Liquid Permanent Red substrate kit.
>The KPL universal blocker Gayle suggested I don't know. However, I am familiar with a similar Dako product called Dual Endogenous Enzyme Block. This blocker also applied before the IHC procedure also effectively destroyed some of my epitopes resulting in a bit too clean staining result indeed.... So be careful with this stuff.  
>Furthermore, I am using PowerVision-AP with great results.
>Hope this helps.
>
>Chris van der Loos, PhD
>Dept. of Pathology
>Academical Medical Center
>Meibergdreef 9
>NL-1105 AZ Amsterdam 
>The Netherlands
>phone: +31 20 5665631  
>
>----- Original Message ----- 
>>From  Ian Montgomery  
>Date  Thu, 18 Nov 2004 17:22:06 +0000 
>To  histonet@lists.utsouthwestern.edu 
>Subject  Fwd: Re: [Histonet] Envision. 
>
>  
>
>>Question:  what buffer are you using?   If you use PBS, then you will have 
>>phosphate ions available for AP, TRIS buffered saline is preferred with AP 
>>protocols.
>>    
>>
>Gayle,
>         I'm using TBS as suggested by Dako. With and without Tween, but 
>still the same result.
>
>
>
>  
>
>>At 08:52 AM 11/18/2004, you wrote:
>>    
>>
>>>        I wonder if anyone can offer any hints and tips for Dako 
>>>Envision. Localisation is fine, HRP is clean but alkaline phosphatase, 
>>>despite my efforts, is still giving me quite a lot of background. This 
>>>manifests itself as a fine red ppt. over the specimen.
>>>Ian.
>>>
>>>Dr. Ian Montgomery,
>>>Histotechnology,
>>>Graham Kerr Building,
>>>Institute of Biomedical & Life Sciences,
>>>University of Glasgow,
>>>Glasgow,
>>>      
>>>
>
>
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>
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