Re: [Histonet] antibodies against phosphorylated proteins

From:"Joanne Mauger"

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"  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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