[Histonet] Re: Immunofluorescent foci moving?

[Gayle Callis]

Vectashield also comes as a hard set.

At 06:44 AM 1/4/2007, you wrote:
>Dear all
>Thanks for your help. I think the problem is that the mounting media is 
>vectashield which remains aqeous and does not hard set, I will change to 
>hard set media, as well as  cetrifuging antibody dilutions and increasing 
>the washes
>Thanks for your help
>Anthony
>
>On 04/01/2007, at 5:36 AM, Edwards, R.E. wrote:
>
>>Could  it  be  that  the  mounting  media  is  incompletely  dry??
>>
>>-----Original Message-----
>>From: 
>><mailto:histonet-bounces <@t> lists.utsouthwestern.edu>histonet-bounces <@t> lists.utsouthwestern.edu
>>[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Anthony
>>joshua
>>Sent: 03 January 2007 03:46
>>To: 
>><mailto:histonet <@t> lists.utsouthwestern.edu>histonet <@t> lists.utsouthwestern.edu
>>Subject: [Histonet] Immunofluorescent foci moving?
>>
>>Hoping for some help here,
>>Im doing FITC staining of some markers of DNA damage foci (gH2Ax,
>>MDC1) in both paraffin and cell lines and find that apart from the
>>nuclear staining pattern that is expected I see that the foci are moving
>>in a kind of brownian motion. Im sure its not bacteria, as they would be
>>dead long ago. Im using a primary and a labeled secondary. Any opinion
>>as to what causes this or what I can do about it? More air-drying?
>>better fixative? I note there was a previous post about this about a
>>year ago but no one replied.
>>Thanks
>>Anthony
>>
>>BSc(Med) MBBS FRACP
>>Fellow in Cellular and Molecular Biology Department of Medical Oncology
>>Princess Margaret Hospital 610 University Avenue
>>M5G2M9 Toronto
>>"Don't worry about the world coming to an end today. It's already
>>tomorrow in Australia." (Charles Schultz)
>>
>>
>>
>>
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>
>
>BSc(Med) MBBS FRACP
>Fellow in Cellular and Molecular Biology
>Department of Medical Oncology
>Princess Margaret Hospital
>610 University Avenue
>M5G2M9 Toronto
>Pager -416-715-2594
>Lab- 416-946-4501 Ext 5012 (Rm 9-717)
>Cell/ Mobile -416-671-1791
>"Don't worry about the world coming to an end today. It's already
>tomorrow in Australia." (Charles Schultz)
>
>
>

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)