[Histonet] antibodies against phosphorylated proteins

[Joanne Mauger]

Heike-

We use tonsil.
Jo

>>> "Heike Grabsch" <H.I.Grabsch <@t> leeds.ac.uk> 06/01/06 11:51 AM >>>
Jo, 
What do you use as positive control tissue, please?
Heike 
> -----Original Message-----
> From: Joanne Mauger [mailto:mauger <@t> email.chop.edu] 
> Sent: 01 June 2006 13:22
> To: Heike Grabsch; histonet <@t> lists.utsouthwestern.edu 
> Subject: Re: [Histonet] antibodies against phosphorylated proteins
> 
> Heike,
> 
> We use H2A.X on human FFPE tissue. The dilution is 1:50 and HIER
with
> citrate buffer, pH 6. We use ABC detection. Works well.
> 
> Jo Mauger
> 
> >>> "Heike Grabsch" <H.I.Grabsch <@t> leeds.ac.uk> 05/31/06 10:32 AM >>>
> I am working with formalin fixed paraffin embedded tissue and trying
> to
> detect phosphorylated proteins by immunohistochemistry. The
antibodies
> are meant to be specific for the phosphorylated protein only.
> The one antibody I am trying to work up in particular is the
> phospho-H2AX from Upstate.
> Is there anybody out there who has an idea whether and if then how
> fixation may effect phosphorylation sides? Can they be lost or
> destroyed
> due to fixation? So the staining could be false negative?
> I hope somebody can comment on this
> 
> Thanks,
> Heike
> 
> Dr Heike Grabsch, MRCPath MD
> Gastrointestinal Research Group
> Pathology and Tumour Biology
> JIF Building, Level 4, Room 4.13
> Leeds Institute for Molecular Medicine
> St James's University Hospital
> Beckett Street
> Leeds
> LS9 7TF
> United Kingdom
> 
> phone 0044 113 3438626
> fax    0044 113 3438431
> 
> 
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