[Histonet] Pms 2 antibody trouble

[Hoekert, W.E.J.]

This is what we are doing on our Benchmark XT's:
 
MLH1: cc1 30', 32' incubation time (Ventana Cell Marque)
MSH2: cc1 60', 60' incubation time (Ventana Cell Marque)
MSH6: cc2 60', 60' incubation time (1:600, Becton Dickinson, clone 44)
PMS2: cc1 30', 32' incubation time + amplification (1:50, Becton Dickinson, clone A16-4)
 
Although our MSH6 stain works, i am not satisfied with the morphology. I guess I should try cc1 30'. 
 
Willem Hoekert
OLVG
Amsterdam
 
 
 

________________________________

Van: histonet-bounces <@t> lists.utsouthwestern.edu namens Amber McKenzie
Verzonden: do 12-1-2012 22:14
Aan: Freeman, Carol
CC: <histonet <@t> lists.utsouthwestern.edu>
Onderwerp: RE: [Histonet] Pms 2 antibody trouble



I just bought the MSI panel antibodies (MLH-1, MSH2, MSH6 and PMS2) from Ventana/Cell Marque and I'm running them on the Ultra and XT: and I'm having trouble, as well, getting them to the desired staining for my pathologists.  I'm staining them at CC1 standard and antibody incubation time is 40 min and I use the amplifier except the MSH2 which is the only one that has been signed off on at 32 min incubation time, no amplifier and mild CC1.  What protocol are you using?

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Karen Lahti
Sent: Thursday, January 12, 2012 3:05 PM
To: Freeman, Carol
Cc: <histonet <@t> lists.utsouthwestern.edu>
Subject: Re: [Histonet] Pms 2 antibody trouble

We use the Biocare concentrated antibody on the Biocare Intellipath. We are diluting at 1:100 and have very good consistent staining. Give me a call if you want to discuss further.

 Karen Lahti, HT, QIHC
Arizona Digestive Health
602-687-7468


On Jan 12, 2012, at 9:42 AM, "Freeman, Carol" <Carol.Freeman <@t> utoledo.edu> wrote:

>
>
> I am curious if anyone is having trouble validating PMS2 antibody for
> the MSI panel.  We have worked this antbody up extensively and we are
> finding that it is on either side of the spectrum just too light (it is
> either light staining if any at all).   We have stained a few cases
> beautifully but nothing is consistent.  We are looking to try another
> vendor's antibodies, but were hoping for some feedback as to which labs
> have found success in this area.  I have never had trouble like this
> with an antibody.  The severe problem here is that with MSI testing, we
> are looking for lack of staining with this antibody to show a positive
> result so light patchy staining leaves a lot of room for error.  Any
> responses are appreciated.
>
> Thank you,
> Carol E. Freeman HTL (ASCP) B.S.
> Department of Pathology
> University of Toledo Medical Center
> 3000 Arlington Avenue
> Toledo, OH 43614-5807
> carol.freeman <@t> utoledo.edu
> (419)383-5639
>
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