[Histonet] RE: CD4 1F6 clone (Steven Weston)

[Path Dynamo]

We use this clone for rhesus macaques FFPE and it works. Not the best but still works.
We do 1:20 dilution,  60' incubation, high pH 9.0 HIER (BondRX or Biocare chamber), and H2O2 block before protein block.
We know it works better on frozen than on FFPE.

I hope this helps.
Best
Pathology Dynamics

<div>-------- Original message --------</div><div>From: histonet-request <@t> lists.utsouthwestern.edu </div><div>Date:09/01/2014  10:00 AM  (GMT-08:00) </div><div>To: histonet <@t> lists.utsouthwestern.edu </div><div>Subject: Histonet Digest, Vol 130, Issue 1 </div><div>
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Today's Topics:

   1. CD4 1F6 clone (Steven Weston)


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Message: 1
Date: Sun, 31 Aug 2014 23:49:20 +0000
From: Steven Weston <Steven.Weston <@t> utas.edu.au>
Subject: [Histonet] CD4 1F6 clone
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <D029F19F.1D09E%Steven.Weston <@t> utas.edu.au>
Content-Type: text/plain; CHARSET=US-ASCII

I also tried it without H2O2 with no result either.
Anyone else had these problems with this clone ???
Interestingly in the past I used the duo pack of CD4/8 that had the 1F6 for cd4 and the 4B11 clone of cd8 and they both worked.
May have to try the 4b12 clone for cd4.
regards
Steve Weston

Message: 10
Date: Sat, 30 Aug 2014 15:44:09 +0530
From: Neeraj Singh <neerajbioxford <@t> gmail.com<mailto:neerajbioxford <@t> gmail.com>>
Subject: Re: [Histonet] Re cd4 1F6 clone
To: Steven Weston <Steven.Weston <@t> utas.edu.au<mailto:Steven.Weston <@t> utas.edu.au>>
Cc: "histonet <@t> lists.utsouthwestern.edu<mailto:histonet <@t> lists.utsouthwestern.edu>"
<histonet <@t> lists.utsouthwestern.edu<mailto:histonet <@t> lists.utsouthwestern.edu>>
Message-ID: <1663FC3E-E152-411F-B553-83D3D426ACEE <@t> gmail.com<mailto:1663FC3E-E152-411F-B553-83D3D426ACEE <@t> gmail.com>>
Content-Type: text/plain; charset=us-ascii

Try with no H2O2 at all for this clone. it will hill you.

Thanks

Neeraj


On Aug 30, 2014, at 1:40 PM, Steven Weston <Steven.Weston <@t> utas.edu.au<mailto:Steven.Weston <@t> utas.edu.au>> wrote:

Hi Histonetters
Has anyone else had difficulty getting  this clone to work? cd4 1F6 from leica microsystems (formerly novocastra).
I've tried just about every trick in my book and still no staining.
Particularly i made sure i did the peroxidase block prior to HIER (i even tried using it after the primary) and have tried low, high and middle pH buffers with antibody concentrations from 1/10-1/40 for 60 and 90 mins and use envision plus detection with DAB plus. I've had two batches of this over the last ten years and neither has worked.
Anyone else with the same experience ?
regards
steve weston
lab manager
Breathe-Well CRE
UTAS-SOM

Steve Weston
Lab Manager
Breathe-Well Centre of Research Excellence for Chronic Respiratory Disease.
UTAS-SOM
0408990859
62264871




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