SPAM-LOW: [Histonet] phosphoantibodies

[Patsy Ruegg]

Here is a fixative we found in a paper on Phospho antibodies when we did a
WS at NSH in 2007.

2.0mM EDTA

2.0mM EGTA

2.0mM Vanadate

40mM NaF

20mMpNPP(para-nitrophenyl phosphate)

We used this fixative compared to routine 10% NBF and did not see any
differences in IHC staining with phospho antibodies.


Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg <@t> ihctech.net
web site www.ihctech.net
 

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of MKing
Sent: Monday, April 19, 2010 8:25 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: SPAM-LOW: [Histonet] phosphoantibodies

Ashley,
Phosphatases are relatively resistant to fixation which is why the 
histochemical phosphatase methods work.  You may want to try putting 
phosphatase inhibitors in you fix or post-fix solutions to stop 
dephosphorylation, and see if it makes a difference in your ihc.  I 
think Sigma has a table of reagents somewhere on their website and I'll 
email the link if I can find it.
Good luck,
Mike
UF Pharmacology & Therapeutics

------------------------
Date: Fri, 16 Apr 2010 14:43:15 -0500
From: "Troutman, Kenneth A" <kenneth.a.troutman <@t> Vanderbilt.Edu>
Subject: [Histonet] Phospho antibodies and fixation
To: "Histonet <@t> lists.utsouthwestern.edu"
	<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:<7B310892042DA74CB3590053F424CFE60B7480EFBC <@t> ITS-HCWNEM06.ds.Vande
rbilt.edu>
	Content-Type: text/plain; charset="iso-8859-1"
Does anyone have any information on phospho antibodies and fixation?...
Thanks,
Ashley Troutman BS, HT(ASCP) QIHC
Vanderbilt University Histopathology
1301 Medical Center Drive TVC 4532
Nashville, TN  37232
615-343-9134

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