[Histonet] RE: Immuno's

[Goins, Tresa]

Valerie -

All of our IHC stains are done manually.  
As an animal tissue lab, our antibody sources will differ, but the best chromagen I have use by far is Biocare's Vulcan Fast Red.  I have tried other sources (cheaper) for the new fuchsin chromagen, but they do not produce the same vivid red stain as Vulcan.

Our secondary antibodies are from KPL (biotinylated anti-mouse or anti-rabbit) with KPL alkaline phosphatase.

Good luck


Tresa Goins
Histopathology Supervisor
Department of Livestock
Bozeman, Montana



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Hannen, Valerie
Sent: Monday, April 11, 2011 8:26 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Immuno's

Hi Folks..
 
Looking for a bit of help. Is there anyone out there who are doing their
immuno's manually?  Our Pathologist has just informed us that our
immuno's are ALL too weak.
He states that they always have been, as he compares them to slides that
we get back from the  reference labs that we use for the antibodies that
we don't do in-house.
 
If you are doing them manually...what companies are you using for
antibodies, detection kit and chromogen? And are your stains really
dark?
 
I informed our Pathologist that I know for a fact that the reference
labs that we use, do the immuno's on stainers, and naturally these
stains will be consistently be stronger.
 
I am trying to see if I can keep costs down by trying not to have to buy
a stainer and continue to do the immuno's manually.
 
If you all know of a wet-workshop or of a immuno. company who would send
someone to our facility to teach us ( the latter is preferred, since my
section chief is going to retire in 1 1/2 weeks, I will be taking over
her duties and this is going to leave us with only myself and one other
tech, until we can get a replacement for my position).
 
Any help or advice that can be given will be greatly appreciated.
 
Thanks so much.
 
Valerie Hannen, MLT(ASCP),HTL,SU (FL)
Parrish Medical Center
Titusville,Florida


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