[Histonet] TTF-1/background staining

[Anthony Reilly]

Hi Paula
 
I do not believe that RTU Abs really exist.  No company can produce an antibody that is suitable for use by all laboratories taking in to consideration how tissue can be handled differently from lab to lab.  I have used many RTU Abs which I have had to dilute to get optimal staining with Cam 5.2 a good example.  On the Bond I was using it at 1:25.  try titreing your Ab and see if that will reduce/eliminate your background.
 
regards
Tony

>>> Debra Siena <DSiena <@t> statlab.com> 4/02/2011 8:02 am >>>
Hi Paula,

I am probably reading between the lines here but on your TTF protocol do you use a streptavidin biotin detection system and if so do you block for endogenous biotin, liver will have endogenous biotin where lung may not have as much?  thanks

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
Direct: 972-436-1010  x229 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Paula Lucas
Sent: Thursday, February 03, 2011 2:38 PM
To: histonet <@t> lists.utsouthwestern.edu 
Subject: [Histonet] TTF-1/background staining

How can I eliminate the background or as you say, non-specific staining?



I'm no expert here, I admit, and so I'm asking for your help and
suggestions.  I have searched the Histonet archives, and a lot of what I'm
seeing deals with animal or rodent tissues, and a lot of it was confusing to
me.



To give you a little background info:

We use the Lab Vision stainer and the TTF-1 antibody we use is from Cell
Marque.  We were having issues with this marker from Lab Vision, so we
switched to TTF-1 a while ago.  We use the UltraVision LP detection kit from
Lab Vision. It's a polymer driven detection kit. 



The antibody is a ready to use, and we have the time set at 30 minutes.  



We were getting the background staining on a liver specimen last week, and
we also had this problem today on a lung case.  My doctor is getting
frustrated, and wants me to do something about this, and to repeat the TTF-1
stain on the lung, so if someone can give me some suggestions to try, I'm
ready to try them.



Thanks in advance,

Paula

Lab Manager

Bio-Path Medical Group

Fountain Valley, CA

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