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 pru...@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of MKing Sent: Monday, April 19, 2010 8:25 AM To: histonet@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.trout...@vanderbilt.edu> Subject: [Histonet] Phospho antibodies and fixation To: "Histonet@lists.utsouthwestern.edu" <Histonet@lists.utsouthwestern.edu> Message-ID:<7b310892042da74cb3590053f424cfe60b7480e...@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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet