[Histonet] Re: Histonet Digest, Vol 104, Issue 15
Madeleine Huey
madeleinehuey <@t> gmail.com
Fri Jul 13 15:31:53 CDT 2012
All,
ER, clone SP1 is availuable in www.epitomics.com (cat. #4200-1) for $205.
Hope this help!
Madeleine Huey
Supervisor - Pathology (IPOX & Histology)
madeleine_h <@t> elcaminohospital.org
On Fri, Jul 13, 2012 at 7:08 AM,
<histonet-request <@t> lists.utsouthwestern.edu> wrote:
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> Today's Topics:
>
> 1. RE: Epic Beaker module (Martha Ward-Pathology)
> 2. Re: Methanol in H2O2 explanation (C.M. van der Loos)
> 3. HT/HTL Full Time Opportunities in New York state! (Brannon Owens)
> 4. ER, Clone SP1 (Leigh York)
> 5. Re: Methanol in H2O2 explanation (Hobbs, Carl)
> 6. Beaker module in EPIC (Paula Sicurello)
> 7. p63 and Biocare Medical (Jmyers1)
> 8. RE: ER, Clone SP1 (Patsy Ruegg)
> 9. Re: ER, Clone SP1 (Kim Donadio)
> 10. RE: ER, Clone SP1 (Morken, Timothy)
> 11. Gamma delta T cell and CD T cell detection on FFPE tissue
> (Kim O'Sullivan)
> 12. RE: ER, Clone SP1 (Patsy Ruegg)
> 13. RE: ER, Clone SP1 (Jonathan Cremer)
> 14. RE: Beaker module in EPIC (Martha Ward-Pathology)
> 15. RE: ER, Clone SP1 (Marsh, Nannette)
> 16. Different Avenues To Become Certified (PATTI NELSON HT(ASCP))
> 17. TRAP staining (Sturm, Mary Therese M)
> 18. RE: TRAP staining (McMahon, Loralee A)
> 19. Re: Re: Methanol in H2O2 explanation (Rene J Buesa)
> 20. Re: Re: Methanol in H2O2 explanation (Rene J Buesa)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Thu, 12 Jul 2012 17:04:09 +0000
> From: Martha Ward-Pathology <mward <@t> wakehealth.edu>
> Subject: RE: [Histonet] Epic Beaker module
> To: Paula Sicurello <patpxs <@t> gmail.com>, HistoNet
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <B2CECB1B6665A4479056478F6DE3C4AB0B5E50E1 <@t> exchdb6.medctr.ad.wfubmc.edu>
>
> Content-Type: text/plain; charset="iso-8859-1"
>
> We would interested in any replies as well.
>
>
>
> Martha Ward, MT (ASCP) QIHC
> Manager
>
> Molecular Diagnostics Lab
> Medical Center Boulevard \ Winston-Salem, NC 27157
> p 336.716.2109 \ f 336.716.5890
> mward <@t> wakehealth.edu
>
>
>
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Paula Sicurello
> Sent: Thursday, July 12, 2012 8:54 AM
> To: HistoNet
> Subject: [Histonet] Epic Beaker module
>
> Good Morning One and All,
>
> Does anyone out there have any experience with the Beaker module of the LIS system EPIC? We have tests that require tables and charts to be inserted into the final report, pathologists need to have a case queue and other items.
>
> If you are using Epic/Beaker- please let me know if your experiences and what if anything have you had Epic build into it for you.
>
> Thanks,
>
> Paula
>
> --
> Paula Sicurello, HTL (ASCP)
> Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710
> P: 919.684.2091
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 2
> Date: Thu, 12 Jul 2012 17:23:39 +0000
> From: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>
> Subject: [Histonet] Re: Methanol in H2O2 explanation
> To: "histonet netserver (histonet <@t> lists.utsouthwestern.edu)"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <BDA3BC4760AFA841926D5FCE9D0C16802BD9E5 <@t> MS-APP-804B.amc.intra>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hi,
>
> There is a paper from 1972 (!) by Streefkerk entitled: "Inhibition of erythrocyte pseudoperoxidase activity by treatment with hydrogen peroxidase following methanol. JHC 30:504. So perhaps the origin of methanol is not that mysterious.
>
> I cannot imagine that methanol is still considered for cryo's when performing IHC: it certainly kill many epitopes!
>
>
>
> Cheers, Chris
>
>
>
> Chris van der Loos
>
> Academic Medical Center
>
> Dept. of Pathology
>
> Amsterdam
>
> The Netherlands
>
>
>
> Date: Wed, 11 Jul 2012 20:22:38 +0200
> From: "Gudrun Lang" <gu.lang <@t> gmx.at>
> Subject: [Histonet] Re: Methanol in H2O2 explanation
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <000f01cd5f92$271cb870$75562950$@gmx.at>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Ha! I've found it. It can be found in Dr. Kiernan's book in the chapter
> enzymehistochemistry - the thing, that an electron-donator is used for
> demonstration.
> And he states, that methanol alone also inhibits peroxidase - perhaps by
> the fixation = denaturation effect on the protein?
>
> In the following publication one can see the equations for the
> peroxidase-reaction and this phrase:
> "In this study, we have used a steady state kinetics approach to demonstrate
> that MPO is irreversibly inactivated by excess H2O2 in the absence of
> reducing substrate."
> The Open Enzyme Inhibition Journal, 2009, 2, 28-35 Mechanism-Based
> Inhibition of Myeloperoxidase by Hydrogen Peroxide: Enhancement of
> Inactivation Rate by Organic Donor Substrates
>
> It's rather exhausting to prove the own statements ;-) puh.
>
> Gudrun
>
>
> De informatie opgenomen in dit bericht kan vertrouwelijk zijn en is uitsluitend bestemd voor de geadresseerde. Indien u dit bericht onterecht ontvangt, wordt u verzocht de inhoud niet te gebruiken en de afzender direct te informeren door het bericht te retourneren. Het Academisch Medisch Centrum is een publiekrechtelijke rechtspersoon in de zin van de W.H.W. (Wet Hoger Onderwijs en Wetenschappelijk Onderzoek) en staat geregistreerd bij de Kamer van Koophandel voor Amsterdam onder nr. 34362777. De Algemene Inkoop Voorwaarden van het AMC zijn van toepassing op en maken integraal onderdeel uit van alle rechtsbetrekkingen, daaronder mede verstaan alle inkoop opdrachten en overeenkomsten, tussen AMC en derden. Deze voorwaarden zijn te raadplegen op www.amc.nl en worden op verzoek toegezonden.
>
> ________________________________
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>
>
> ------------------------------
>
> Message: 3
> Date: Thu, 12 Jul 2012 14:48:21 -0400
> From: Brannon Owens <brannon <@t> alliedsearchpartners.com>
> Subject: [Histonet] HT/HTL Full Time Opportunities in New York state!
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <CC249435.176FE%brannon <@t> alliedsearchpartners.com>
> Content-Type: text/plain; charset="US-ASCII"
>
> Allied Search Partners is working with different laboratories in the state
> of New York, looking for qualified and licensed Histotechnologists or
> Histotechnicians.
>
> To Apply Please email or fax resume to Brannon <@t> alliedsearchpartners.com
> <mailto:Brannon <@t> alliedsearchpartners.com> or fax to 888 388 7572. No other
> information is given about location or the organization at this time. Please
> send resume for review by our recruiters and all qualified candidates will
> be submitted to HR for further review. Once we have had time to further
> consider your application, then more information will be made available.
> Thank you!
>
> Position 1:
> Immunohistochemistry Technician/Technologist (IHC)
> Schedule: Tuesday-Saturday 2pm-10:30pm
> Location: Port Chester, NY
> Requirements: experience in IHC, a NY license, ASCP HT/HTL certification
>
> Position 2:
> Histotechnician or Histotechnologist
> Schedule: Tuesday-Saturday 7am-3:30pm
> Location: Port Chester, NY
> Requirements: NY Licensed, ASCP HT/HTL certification
>
> Position 3:
> Histotechnician or Histotechnologist
> Schedule: Full time/Permanent Day Shift
> Location: Binghamton, NY
> Requirements: NY Licensed, ASCP HT/HTL certification
>
> --
> Brannon Owens
> Recruitment Manager
> Allied Search Partners
>
>
> T: 888.388.7571 ext. 106
>
> F: 888.388.7572
>
>
>
> To view a complete list of Allied Search Partners current openings go to:
> http://www.alliedsearchpartners.com/careers.php
>
> LinkedIn: http://www.linkedin.com/pub/brannon-owens/28/528/823
>
>
>
>
>
>
>
> ------------------------------
>
> Message: 4
> Date: Thu, 12 Jul 2012 15:06:27 -0400
> From: "Leigh York" <lyork <@t> kwbpathology.com>
> Subject: [Histonet] ER, Clone SP1
> To: <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <B422B89C676A4B2F8895D2C4AF3AAE1C <@t> KWBPA.local>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Dako is going to discontinue the ER , SP1 clone. We really want to use the SP1 clone, but do not want to use an ASR. We can get it from Ventana only we are told. Can anyone elaborate on how they are handling this in their lab or if they have switched to an ASR?
>
> Thanks,
>
> Leigh York
>
> ------------------------------
>
> Message: 5
> Date: Thu, 12 Jul 2012 20:27:37 +0100
> From: "Hobbs, Carl" <carl.hobbs <@t> kcl.ac.uk>
> Subject: [Histonet] Re: Methanol in H2O2 explanation
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <11D9615B89C10747B1C985966A63D7CA38626A5736 <@t> KCL-MAIL04.kclad.ds.kcl.ac.uk>
>
> Content-Type: text/plain; charset="us-ascii"
>
> "H202 mixed with Buffer here.(instead of aq.) That's OK too, right?"
>
> Right.
>
> An addition to the misconceptions that Tim interestingly elucidates: some use H2O2 in buffer because they think that , as the enzyme is performing a physiologic catalytic reaction, it should be at a physiological pH, in a physiological buffer.
> Well....if it's a frozen section, it's been fixed; if it's a P.wax section, it's been fixed in Formalin, fixed in alcohol, subjected to 60C heat.....so, the fact that endog. Peroxidases survive those assaults, indicates that physiologic conditions no longer apply?
>
> Re use of methanol/ethanol....I use IMS. Always have done.
> NB: great excess of substrate can " kill " enzymes ( irreversible substrate inhibition): that is why we add a great excess of H2O2 ( substrate) to "kill" endog. Pxs.
> We then use a stoichiometric quantity of the same reagent ( H2O2) in the presence of DAB/AEC, to visualize sites of AB:Ag interaction.
>
> "For the enzymatic activity of peroxidase it needs an electron-donator"
> The enzymatic activity of peroxidase needs no electron donor, as regards Immuno.....it will "digest" H2O2 into water and oxygen gas.
> However, if you wish to use HRP as a reporter molecule, you must also add a suitable chromogen ( DAB, AEC) that
>
>
> Carl Hobbs
> Histology Manager
> Wolfson CARD
> School of Biomedical Sciences
> Kings College London
> Guys Campus
> SE1 1UL
> Tel: 020 78486813
> Fax: 020 78486816
> 020 78486813
>
>
>
>
> ------------------------------
>
> Message: 6
> Date: Thu, 12 Jul 2012 16:04:51 -0400
> From: Paula Sicurello <patpxs <@t> gmail.com>
> Subject: [Histonet] Beaker module in EPIC
> To: HistoNet <histonet <@t> lists.utsouthwestern.edu>,
> Microscopy <@t> microscopy.com
> Message-ID:
> <CAPSjddYA=3LUtYM_LA9YmQm1=4St9CjRPCC3Mg8Q=CJOA-aHgw <@t> mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
>
> Hello all Listers,
>
> I've already asked the Histonet but I'm wondering about a different thing
> this time.
>
> Does anyone have experience with Beaker in conjunction with Anatomic
> Pathology, Surgical Pathology or the multiple other clinical labs that are
> in hospitals today?
>
> I work in Anatomic/Surgical Pathology which encompasses all the Clinical
> Laboratory services here at Duke. We have been told to determine what we
> need for the Beaker module to be useful to us. I've seen a brief demo. as
> to how it will work for the ordering clinicians (they can order every test
> STAT!).
>
> Please, send me any of the pluses or minuses that you've experienced using
> Beaker.
>
> Thanks,
>
> Paula :-)
>
> --
> Paula Sicurello, HTL (ASCP)
> Supervisor, Clinical Electron Microscopy Laboratory
> Duke University Health System
> Rm.#251M, Duke South, Green Zone
> Durham, North Carolina 27710
> P: 919.684.2091
>
>
> ------------------------------
>
> Message: 7
> Date: Thu, 12 Jul 2012 17:19:35 -0400 (EDT)
> From: Jmyers1 <jmyers1 <@t> aol.com>
> Subject: [Histonet] p63 and Biocare Medical
> To: histonet <@t> lists.utsouthwestern.edu, jmyers <@t> biocare.net
> Message-ID: <8CF2E8B2C0332D7-7DC-410D <@t> webmail-d128.sysops.aol.com>
> Content-Type: text/plain; charset="utf-8"
>
>
> Fellow Histonetters:
> Over the past few days there has been some discussion on this forum regarding Biocare Medical's ability to continue offering the p63 antibody, some of which has been correct and some of it incorrect. Please be assured that Biocare still has the right to sell products that contain p63, but future U.S. sales rights are being negotiated and are subject to change. At present, only Biocare and Ventana Medical Systems are licensed to distribute IVD-labeled products that contain p63. If you have any concerns about this situation, please direct them to the Biocare sales representative that is responsible for the area in which your laboratory is located.
> Sincerely,
> Joe Myers, M.S., CT(ASCP)QIHC
> Senior Technical Sales Specialist – Biocare Medical, LLC
> (and Chair, IHC Resource Group – National Society for Histotechnology)
>
>
> ------------------------------
>
> Message: 8
> Date: Thu, 12 Jul 2012 15:45:20 -0600
> From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
> Subject: RE: [Histonet] ER, Clone SP1
> To: "'Leigh York'" <lyork <@t> kwbpathology.com>,
> <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <6959269F7DA142FCB3EEEC3CFA29B6E2 <@t> Patsyoffice>
> Content-Type: text/plain; charset="us-ascii"
>
> >From what I understand the SP1 Er clone has also been discontinued in USA by
> Leica/Novacastra, I am told it is still being sold by Leica in Europe? Must
> be another licensing issue because some are soooo greedy. I knew this was
> going to happen and stock piled some of the ER SP1, but it won't last
> forever.
>
> Patsy Ruegg, HT(ASCP)QIHC
> Ruegg Consulting
> 40864 Arkansas Ave
> Bennett, CO 80102
> Phone: 303-644-4538
> Fax: 720-859-4110
> pruegg <@t> ihctech.net
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Leigh York
> Sent: Thursday, July 12, 2012 1:06 PM
> To: Histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] ER, Clone SP1
>
> Dako is going to discontinue the ER , SP1 clone. We really want to use the
> SP1 clone, but do not want to use an ASR. We can get it from Ventana only
> we are told. Can anyone elaborate on how they are handling this in their
> lab or if they have switched to an ASR?
>
> Thanks,
>
> Leigh York
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 9
> Date: Thu, 12 Jul 2012 17:54:31 -0400
> From: Kim Donadio <one_angel_secret <@t> yahoo.com>
> Subject: Re: [Histonet] ER, Clone SP1
> To: Patsy Ruegg <pruegg <@t> ihctech.net>
> Cc: "<Histonet <@t> lists.utsouthwestern.edu>"
> <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <876C9001-87DB-4723-8F71-D99F14D303EA <@t> yahoo.com>
> Content-Type: text/plain; charset=us-ascii
>
> I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue?
>
> Sent from my iPhone
>
> On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" <pruegg <@t> ihctech.net> wrote:
>
>>> From what I understand the SP1 Er clone has also been discontinued in USA by
>> Leica/Novacastra, I am told it is still being sold by Leica in Europe? Must
>> be another licensing issue because some are soooo greedy. I knew this was
>> going to happen and stock piled some of the ER SP1, but it won't last
>> forever.
>>
>> Patsy Ruegg, HT(ASCP)QIHC
>> Ruegg Consulting
>> 40864 Arkansas Ave
>> Bennett, CO 80102
>> Phone: 303-644-4538
>> Fax: 720-859-4110
>> pruegg <@t> ihctech.net
>>
>> -----Original Message-----
>> From: histonet-bounces <@t> lists.utsouthwestern.edu
>> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Leigh York
>> Sent: Thursday, July 12, 2012 1:06 PM
>> To: Histonet <@t> lists.utsouthwestern.edu
>> Subject: [Histonet] ER, Clone SP1
>>
>> Dako is going to discontinue the ER , SP1 clone. We really want to use the
>> SP1 clone, but do not want to use an ASR. We can get it from Ventana only
>> we are told. Can anyone elaborate on how they are handling this in their
>> lab or if they have switched to an ASR?
>>
>> Thanks,
>>
>> Leigh York
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 10
> Date: Thu, 12 Jul 2012 15:05:17 -0700
> From: "Morken, Timothy" <Timothy.Morken <@t> ucsfmedctr.org>
> Subject: RE: [Histonet] ER, Clone SP1
> To: "Kim Donadio" <one_angel_secret <@t> yahoo.com>, "Patsy Ruegg"
> <pruegg <@t> ihctech.net>
> Cc: "<Histonet <@t> lists.utsouthwestern.edu>"
> <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <8D7C2D242DBD45498006B21122072BF8B5BBAC03 <@t> MCINFRWEM003.ucsfmedicalcenter.org>
>
> Content-Type: text/plain; charset=us-ascii
>
> It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues.
>
> For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company.
>
> Tim Morken
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim Donadio
> Sent: Thursday, July 12, 2012 2:55 PM
> To: Patsy Ruegg
> Cc: <Histonet <@t> lists.utsouthwestern.edu>
> Subject: Re: [Histonet] ER, Clone SP1
>
> I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue?
>
> Sent from my iPhone
>
> On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" <pruegg <@t> ihctech.net> wrote:
>
>>> From what I understand the SP1 Er clone has also been discontinued in
>>> USA by
>> Leica/Novacastra, I am told it is still being sold by Leica in Europe?
>> Must be another licensing issue because some are soooo greedy. I knew
>> this was going to happen and stock piled some of the ER SP1, but it
>> won't last forever.
>>
>> Patsy Ruegg, HT(ASCP)QIHC
>> Ruegg Consulting
>> 40864 Arkansas Ave
>> Bennett, CO 80102
>> Phone: 303-644-4538
>> Fax: 720-859-4110
>> pruegg <@t> ihctech.net
>>
>> -----Original Message-----
>> From: histonet-bounces <@t> lists.utsouthwestern.edu
>> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Leigh
>> York
>> Sent: Thursday, July 12, 2012 1:06 PM
>> To: Histonet <@t> lists.utsouthwestern.edu
>> Subject: [Histonet] ER, Clone SP1
>>
>> Dako is going to discontinue the ER , SP1 clone. We really want to
>> use the
>> SP1 clone, but do not want to use an ASR. We can get it from Ventana
>> only we are told. Can anyone elaborate on how they are handling this
>> in their lab or if they have switched to an ASR?
>>
>> Thanks,
>>
>> Leigh York
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 11
> Date: Fri, 13 Jul 2012 09:20:25 +1000
> From: "Kim O'Sullivan" <kim.osullivan <@t> monash.edu>
> Subject: [Histonet] Gamma delta T cell and CD T cell detection on FFPE
> tissue
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
> <CALJ_c9-iXahtW1nTrG2auW8r_U8eVmS=ti-+uu-dmrrfGFwTbw <@t> mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
>
> Hi everyone,
>
> Can anyone tell me of a commercially available antibody that works on FFPE
> human tissue for gamma delta T cells ( TCR)? Secondly does anyone have a
> protocol for CD4 T cells on human tissue, I have tried both DAKO mouse anti
> human and both clones available from Novacastra (4B12 and 1F6) using
> microwave antigen retrieval (citrate buffer pH 6, EDTA pH 8 and TRIS HCL pH
> 9) and pressure cooker with the same buffers for different time periods (
> 1, 2,3,4,and 5 minutes). Any suggestions, protocols or alternative
> antibodies would be appreciated!
>
> Regards
>
>
> ------------------------------
>
> Message: 12
> Date: Thu, 12 Jul 2012 20:44:33 -0600
> From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
> Subject: RE: [Histonet] ER, Clone SP1
> To: "'Morken, Timothy'" <Timothy.Morken <@t> ucsfmedctr.org>, "'Kim
> Donadio'" <one_angel_secret <@t> yahoo.com>
> Cc: Histonet <@t> lists.utsouthwestern.edu
> Message-ID: <0227518F256543848B5009F06BDF4EEB <@t> Patsyoffice>
> Content-Type: text/plain; charset="us-ascii"
>
> Yea you are right Tim, I just wonder what ever happened to trying to get the
> best ab for the patient, SP1 is in my opinion the best ER ab but now the
> patient suffers because we can't use what is best for them, there is
> something wrong with that picture.
>
> Patsy Ruegg, HT(ASCP)QIHC
> Ruegg Consulting
> 40864 Arkansas Ave
> Bennett, CO 80102
> Phone: 303-644-4538
> Fax: 720-859-4110
> pruegg <@t> ihctech.net
>
> -----Original Message-----
> From: Morken, Timothy [mailto:Timothy.Morken <@t> ucsfmedctr.org]
> Sent: Thursday, July 12, 2012 4:05 PM
> To: Kim Donadio; Patsy Ruegg
> Cc: <Histonet <@t> lists.utsouthwestern.edu>
> Subject: RE: [Histonet] ER, Clone SP1
>
> It is most likely licensing issues between companies. A company may carry it
> under license from another company but the terms change and the licensee
> either loses the license (the other company won't license it), can't afford
> it or other issues.
>
> For example, when I worked in the antibody industry we had several
> antibodies for a certain target but another company got a patent on the
> target and all antibodies to the target (still not sure why that is
> possible!). They finally decided to enforce their patent and wanted a
> licensing fee that was higher than our annual sales of all the antibodies
> put together. Obviously not a viable deal for our company.
>
> Tim Morken
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim Donadio
> Sent: Thursday, July 12, 2012 2:55 PM
> To: Patsy Ruegg
> Cc: <Histonet <@t> lists.utsouthwestern.edu>
> Subject: Re: [Histonet] ER, Clone SP1
>
> I'm curious as to why all if a sudden some of these can be sold else where
> and not in the USA. Is it a FDA issue?
>
> Sent from my iPhone
>
> On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" <pruegg <@t> ihctech.net> wrote:
>
>>> From what I understand the SP1 Er clone has also been discontinued in
>>> USA by
>> Leica/Novacastra, I am told it is still being sold by Leica in Europe?
>> Must be another licensing issue because some are soooo greedy. I knew
>> this was going to happen and stock piled some of the ER SP1, but it
>> won't last forever.
>>
>> Patsy Ruegg, HT(ASCP)QIHC
>> Ruegg Consulting
>> 40864 Arkansas Ave
>> Bennett, CO 80102
>> Phone: 303-644-4538
>> Fax: 720-859-4110
>> pruegg <@t> ihctech.net
>>
>> -----Original Message-----
>> From: histonet-bounces <@t> lists.utsouthwestern.edu
>> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Leigh
>> York
>> Sent: Thursday, July 12, 2012 1:06 PM
>> To: Histonet <@t> lists.utsouthwestern.edu
>> Subject: [Histonet] ER, Clone SP1
>>
>> Dako is going to discontinue the ER , SP1 clone. We really want to
>> use the
>> SP1 clone, but do not want to use an ASR. We can get it from Ventana
>> only we are told. Can anyone elaborate on how they are handling this
>> in their lab or if they have switched to an ASR?
>>
>> Thanks,
>>
>> Leigh York
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
>
> ------------------------------
>
> Message: 13
> Date: Fri, 13 Jul 2012 06:45:20 +0000
> From: Jonathan Cremer <Jonathan.Cremer <@t> med.kuleuven.be>
> Subject: RE: [Histonet] ER, Clone SP1
> To: "Morken, Timothy" <Timothy.Morken <@t> ucsfmedctr.org>, Kim Donadio
> <one_angel_secret <@t> yahoo.com>, Patsy Ruegg <pruegg <@t> ihctech.net>
> Cc: "<Histonet <@t> lists.utsouthwestern.edu>"
> <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <C35C2709C22FC2458D6A918FACC338A30FA3087E <@t> ICTS-S-MBX5.luna.kuleuven.be>
>
> Content-Type: text/plain; charset="us-ascii"
>
> That drives me up the wall as well. How can you possibly patent something that has been made by nature, and in theory is present in every single human being (I'm talking about any random gene here)? That would be like Newton patenting gravity and then going around and charging everyone a licensing fee for staying on earth instead of floating off in space, or claiming every apple that has fallen off a tree.
> It's ridiculous!
> ---
>
>
>
> ________________________________________
> Van: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] namens Morken, Timothy [Timothy.Morken <@t> ucsfmedctr.org]
> Verzonden: vrijdag 13 juli 2012 0:05
> To: Kim Donadio; Patsy Ruegg
> Cc: <Histonet <@t> lists.utsouthwestern.edu>
> Onderwerp: RE: [Histonet] ER, Clone SP1
>
> It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues.
>
> For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company.
>
> Tim Morken
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim Donadio
> Sent: Thursday, July 12, 2012 2:55 PM
> To: Patsy Ruegg
> Cc: <Histonet <@t> lists.utsouthwestern.edu>
> Subject: Re: [Histonet] ER, Clone SP1
>
> I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue?
>
> Sent from my iPhone
>
> On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" <pruegg <@t> ihctech.net> wrote:
>
>>> From what I understand the SP1 Er clone has also been discontinued in
>>> USA by
>> Leica/Novacastra, I am told it is still being sold by Leica in Europe?
>> Must be another licensing issue because some are soooo greedy. I knew
>> this was going to happen and stock piled some of the ER SP1, but it
>> won't last forever.
>>
>> Patsy Ruegg, HT(ASCP)QIHC
>> Ruegg Consulting
>> 40864 Arkansas Ave
>> Bennett, CO 80102
>> Phone: 303-644-4538
>> Fax: 720-859-4110
>> pruegg <@t> ihctech.net
>>
>> -----Original Message-----
>> From: histonet-bounces <@t> lists.utsouthwestern.edu
>> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Leigh
>> York
>> Sent: Thursday, July 12, 2012 1:06 PM
>> To: Histonet <@t> lists.utsouthwestern.edu
>> Subject: [Histonet] ER, Clone SP1
>>
>> Dako is going to discontinue the ER , SP1 clone. We really want to
>> use the
>> SP1 clone, but do not want to use an ASR. We can get it from Ventana
>> only we are told. Can anyone elaborate on how they are handling this
>> in their lab or if they have switched to an ASR?
>>
>> Thanks,
>>
>> Leigh York
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 14
> Date: Fri, 13 Jul 2012 11:42:01 +0000
> From: Martha Ward-Pathology <mward <@t> wakehealth.edu>
> Subject: RE: [Histonet] Beaker module in EPIC
> To: Paula Sicurello <patpxs <@t> gmail.com>, HistoNet
> <histonet <@t> lists.utsouthwestern.edu>, "Microscopy <@t> microscopy.com"
> <Microscopy <@t> microscopy.com>
> Message-ID:
> <B2CECB1B6665A4479056478F6DE3C4AB0B5E5246 <@t> exchdb6.medctr.ad.wfubmc.edu>
>
> Content-Type: text/plain; charset="us-ascii"
>
> Again, we would be interested in any replies as well. Our institution is going live with Epic this fall but the labs will not be using Beaker until later on. We are still trying to figure out how all this will work and welcome all information.
> Paula, how soon will Duke be up and using Beaker?
>
> Martha Ward
> Wake Forest Baptist Health
> 336-716-2109
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Paula Sicurello
> Sent: Thursday, July 12, 2012 4:05 PM
> To: HistoNet; Microscopy <@t> microscopy.com
> Subject: [Histonet] Beaker module in EPIC
>
> Hello all Listers,
>
> I've already asked the Histonet but I'm wondering about a different thing this time.
>
> Does anyone have experience with Beaker in conjunction with Anatomic Pathology, Surgical Pathology or the multiple other clinical labs that are in hospitals today?
>
> I work in Anatomic/Surgical Pathology which encompasses all the Clinical Laboratory services here at Duke. We have been told to determine what we need for the Beaker module to be useful to us. I've seen a brief demo. as to how it will work for the ordering clinicians (they can order every test STAT!).
>
> Please, send me any of the pluses or minuses that you've experienced using Beaker.
>
> Thanks,
>
> Paula :-)
>
> --
> Paula Sicurello, HTL (ASCP)
> Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710
> P: 919.684.2091
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 15
> Date: Fri, 13 Jul 2012 07:06:34 -0500
> From: "Marsh, Nannette" <NMP <@t> stowers.org>
> Subject: RE: [Histonet] ER, Clone SP1
> To: "'Jonathan Cremer'" <Jonathan.Cremer <@t> med.kuleuven.be>, "Morken,
> Timothy" <Timothy.Morken <@t> ucsfmedctr.org>, Kim Donadio
> <one_angel_secret <@t> yahoo.com>, Patsy Ruegg <pruegg <@t> ihctech.net>
> Cc: "<Histonet <@t> lists.utsouthwestern.edu>"
> <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <2C40E43D1F7A56408C4463FD245DDDF9AE46A699 <@t> EXCHMB-02.stowers-institute.org>
>
> Content-Type: text/plain; charset="us-ascii"
>
> I love the comparison :-)
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jonathan Cremer
> Sent: Friday, July 13, 2012 1:45 AM
> To: Morken, Timothy; Kim Donadio; Patsy Ruegg
> Cc: <Histonet <@t> lists.utsouthwestern.edu>
> Subject: RE: [Histonet] ER, Clone SP1
>
> That drives me up the wall as well. How can you possibly patent something that has been made by nature, and in theory is present in every single human being (I'm talking about any random gene here)? That would be like Newton patenting gravity and then going around and charging everyone a licensing fee for staying on earth instead of floating off in space, or claiming every apple that has fallen off a tree.
> It's ridiculous!
> ---
>
>
>
> ________________________________________
> Van: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] namens Morken, Timothy [Timothy.Morken <@t> ucsfmedctr.org]
> Verzonden: vrijdag 13 juli 2012 0:05
> To: Kim Donadio; Patsy Ruegg
> Cc: <Histonet <@t> lists.utsouthwestern.edu>
> Onderwerp: RE: [Histonet] ER, Clone SP1
>
> It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues.
>
> For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company.
>
> Tim Morken
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim Donadio
> Sent: Thursday, July 12, 2012 2:55 PM
> To: Patsy Ruegg
> Cc: <Histonet <@t> lists.utsouthwestern.edu>
> Subject: Re: [Histonet] ER, Clone SP1
>
> I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue?
>
> Sent from my iPhone
>
> On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" <pruegg <@t> ihctech.net> wrote:
>
>>> From what I understand the SP1 Er clone has also been discontinued in
>>> USA by
>> Leica/Novacastra, I am told it is still being sold by Leica in Europe?
>> Must be another licensing issue because some are soooo greedy. I knew
>> this was going to happen and stock piled some of the ER SP1, but it
>> won't last forever.
>>
>> Patsy Ruegg, HT(ASCP)QIHC
>> Ruegg Consulting
>> 40864 Arkansas Ave
>> Bennett, CO 80102
>> Phone: 303-644-4538
>> Fax: 720-859-4110
>> pruegg <@t> ihctech.net
>>
>> -----Original Message-----
>> From: histonet-bounces <@t> lists.utsouthwestern.edu
>> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Leigh
>> York
>> Sent: Thursday, July 12, 2012 1:06 PM
>> To: Histonet <@t> lists.utsouthwestern.edu
>> Subject: [Histonet] ER, Clone SP1
>>
>> Dako is going to discontinue the ER , SP1 clone. We really want to
>> use the
>> SP1 clone, but do not want to use an ASR. We can get it from Ventana
>> only we are told. Can anyone elaborate on how they are handling this
>> in their lab or if they have switched to an ASR?
>>
>> Thanks,
>>
>> Leigh York
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 16
> Date: Fri, 13 Jul 2012 05:19:18 -0700 (PDT)
> From: "PATTI NELSON HT\(ASCP\)" <nelsonrnch <@t> verizon.net>
> Subject: [Histonet] Different Avenues To Become Certified
> To: Histonet <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <1342181958.20737.YahooMailNeo <@t> web84511.mail.ne1.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Good morning Histo-Land,
>
> A question was asked to me by my Medical Director the other day, so I ask for everyones expert answers to this question.
> Is there any other entities other than ASCP to be certified by for HT/HTL ?
>
> I thank everyone in advance. Have a great weekend.
>
> BEST REGARDS,
>
> PATTI RUBEN-NELSON H.T.(ASCP)
> PNP LABORATORY CONSULTANTS
> SUPERVISOR/DGC
> P.O. BOX 412
> CABAZON, CA. 92230
> cell (909) 841-9761
> nelsonrnch <@t> verizon.net
>
>
> CONFIDENTIALITY NOTICE:
> This message and any included attachments are from Patti Nelson, PNP Laboratory Consultants
> and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or exemption from disclosure under applicable
> law. Unauthorized forwarding, printing, copying, distribution, or use of
> such information is strictly prohibited and may be unlawful. If you are not
> the addressee, please promptly delete this message and notify the sender of
> the delivery error by e-mail or you may call 909-841-9761.
>
> ------------------------------
>
> Message: 17
> Date: Fri, 13 Jul 2012 13:18:06 +0000
> From: "Sturm, Mary Therese M" <mary-sturm <@t> uiowa.edu>
> Subject: [Histonet] TRAP staining
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <17B53F9D0552174BAB6F33E0B69BBD9910847B70 <@t> hc-mailboxc1-n3.healthcare.uiowa.edu>
>
> Content-Type: text/plain; charset="us-ascii"
>
> Currently we are trying to use Sigma Kit 386A for Trap staining of FFPE rabbit spine.....and getting no staining , only tons of gold background. Anyone have experience with this procedure?
>
> Thanks
> Mary Sturm
>
>
>
>
> ________________________________
> Notice: This UI Health Care e-mail (including attachments) is covered by the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is confidential and may be legally privileged. If you are not the intended recipient, you are hereby notified that any retention, dissemination, distribution, or copying of this communication is strictly prohibited. Please reply to the sender that you have received the message in error, then delete it. Thank you.
> ________________________________
>
>
> ------------------------------
>
> Message: 18
> Date: Fri, 13 Jul 2012 09:33:23 -0400
> From: "McMahon, Loralee A" <Loralee_Mcmahon <@t> URMC.Rochester.edu>
> Subject: [Histonet] RE: TRAP staining
> To: "Sturm, Mary Therese M" <mary-sturm <@t> uiowa.edu>,
> "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <EC3287D73321A14799BB96082DE57B4753866373 <@t> URMCMS2.urmc-sh.rochester.edu>
>
> Content-Type: text/plain; charset="us-ascii"
>
> What was your tissue decaled in?
>
> Loralee McMahon, HTL (ASCP)
> Immunohistochemistry Supervisor
> Strong Memorial Hospital
> Department of Surgical Pathology
> (585) 275-7210
> ________________________________________
> From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sturm, Mary Therese M [mary-sturm <@t> uiowa.edu]
> Sent: Friday, July 13, 2012 9:18 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] TRAP staining
>
> Currently we are trying to use Sigma Kit 386A for Trap staining of FFPE rabbit spine.....and getting no staining , only tons of gold background. Anyone have experience with this procedure?
>
> Thanks
> Mary Sturm
>
>
>
>
> ________________________________
> Notice: This UI Health Care e-mail (including attachments) is covered by the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is confidential and may be legally privileged. If you are not the intended recipient, you are hereby notified that any retention, dissemination, distribution, or copying of this communication is strictly prohibited. Please reply to the sender that you have received the message in error, then delete it. Thank you.
> ________________________________
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ------------------------------
>
> Message: 19
> Date: Fri, 13 Jul 2012 06:55:15 -0700 (PDT)
> From: Rene J Buesa <rjbuesa <@t> yahoo.com>
> Subject: Re: [Histonet] Re: Methanol in H2O2 explanation
> To: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>, "histonet
> netserver \(histonet <@t> lists.utsouthwestern.edu\)"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <1342187715.62239.YahooMailNeo <@t> web121405.mail.ne1.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> This is the real explanation.
> Mind that the title refers to erythrocyte pseudoperoxidase inhibition and in the title of the article is the procedural sequence:
> 1- fix the blood smear with methanol, which is the standard procedure, and
> 2- apply H2O2 to quench the peroxidase activity.
> René J.
>
>
> ________________________________
> From: C.M. van der Loos <c.m.vanderloos <@t> amc.uva.nl>
> To: "histonet netserver (histonet <@t> lists.utsouthwestern.edu)" <histonet <@t> lists.utsouthwestern.edu>
> Sent: Thursday, July 12, 2012 1:23 PM
> Subject: [Histonet] Re: Methanol in H2O2 explanation
>
> Hi,
>
> There is a paper from 1972 (!) by Streefkerk entitled: "Inhibition of erythrocyte pseudoperoxidase activity by treatment with hydrogen peroxidase following methanol. JHC 30:504. So perhaps the origin of methanol is not that mysterious.
>
> I cannot imagine that methanol is still considered for cryo's when performing IHC: it certainly kill many epitopes!
>
>
>
> Cheers, Chris
>
>
>
> Chris van der Loos
>
> Academic Medical Center
>
> Dept. of Pathology
>
> Amsterdam
>
> The Netherlands
>
>
>
> Date: Wed, 11 Jul 2012 20:22:38 +0200
> From: "Gudrun Lang" <gu.lang <@t> gmx.at>
> Subject: [Histonet] Re: Methanol in H2O2 explanation
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <000f01cd5f92$271cb870$75562950$@gmx.at>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Ha! I've found it. It can be found in Dr. Kiernan's book in the chapter
> enzymehistochemistry - the thing, that an electron-donator is used for
> demonstration.
> And he states, that methanol alone also inhibits peroxidase - perhaps by
> the fixation = denaturation effect on the protein?
>
> In the following publication one can see the equations for the
> peroxidase-reaction and this phrase:
> "In this study, we have used a steady state kinetics approach to demonstrate
> that MPO is irreversibly inactivated by excess H2O2 in the absence of
> reducing substrate."
> The Open Enzyme Inhibition Journal, 2009, 2, 28-35 Mechanism-Based
> Inhibition of Myeloperoxidase by Hydrogen Peroxide: Enhancement of
> Inactivation Rate by Organic Donor Substrates
>
> It's rather exhausting to prove the own statements ;-) puh.
>
> Gudrun
>
>
> De informatie opgenomen in dit bericht kan vertrouwelijk zijn en is uitsluitend bestemd voor de geadresseerde. Indien u dit bericht onterecht ontvangt, wordt u verzocht de inhoud niet te gebruiken en de afzender direct te informeren door het bericht te retourneren. Het Academisch Medisch Centrum is een publiekrechtelijke rechtspersoon in de zin van de W.H.W. (Wet Hoger Onderwijs en Wetenschappelijk Onderzoek) en staat geregistreerd bij de Kamer van Koophandel voor Amsterdam onder nr. 34362777. De Algemene Inkoop Voorwaarden van het AMC zijn van toepassing op en maken integraal onderdeel uit van alle rechtsbetrekkingen, daaronder mede verstaan alle inkoop opdrachten en overeenkomsten, tussen AMC en derden. Deze voorwaarden zijn te raadplegen op http://www.amc.nl/ en worden op verzoek toegezonden.
>
> ________________________________
>
> This message may contain confidential information and is intended exclusively for the addressee. If you receive this message unintentionally, please do not use the contents but notify the sender immediately by return e-mail. Academic Medical Center is a legal person by public law and is registered at the Chamber of Commerce for Amsterdam under no. 34362777. The AMC General Purchase Terms constitute an integral part of all legal relations, including but not limited to all purchase orders and contracts, between the AMC and third parties. These terms can be downloaded at the website www.amc.nl and will be sent to you at your request.
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> ------------------------------
>
> Message: 20
> Date: Fri, 13 Jul 2012 07:07:42 -0700 (PDT)
> From: Rene J Buesa <rjbuesa <@t> yahoo.com>
> Subject: Re: [Histonet] Re: Methanol in H2O2 explanation
> To: "Hobbs, Carl" <carl.hobbs <@t> kcl.ac.uk>,
> "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
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> I have been fascinated by this long discussion about H2O2 and its use in IHC procedures.
> We have to start from the fact that ALL cells contain peroxidase because ALL cells respire and ALL cells need to use oxygen they receive from blood (or hemolymph in crustaceans and other arthropods).
> The initial IHC procedure were peroxidase-anti-peroxidase (PAP) methods (and many remain as such).
> A fixed cell = a dead cell BUT in spite of being dead, the peroxidase they contain is still able of reacting with the PAP complex.
> The cell, being dead, cannot make more peroxidase, but there is still peroxidase in it after any type of fixation, even with NBF, as Carl points out.
> IF that peroxidase is not "quenched" = INHIBITED the use of a PAP IHC procedure on that uninhibited peroxidase = excessive background in the section after the IHC method is completed.
> So, how to avoid that background? By "quenching" or inhibiting the peroxidase contained in the cell.
> How? With a substance rich in oxygen that will inactivate the peroxidase by "spending" it and that substance is H2O2 used at the start of the IHC protocol.
> René J.
>
>
> ________________________________
> From: "Hobbs, Carl" <carl.hobbs <@t> kcl.ac.uk>
> To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
> Sent: Thursday, July 12, 2012 3:27 PM
> Subject: [Histonet] Re: Methanol in H2O2 explanation
>
> "H202 mixed with Buffer here.(instead of aq.) That's OK too, right?"
>
> Right.
>
> An addition to the misconceptions that Tim interestingly elucidates: some use H2O2 in buffer because they think that , as the enzyme is performing a physiologic catalytic reaction, it should be at a physiological pH, in a physiological buffer.
> Well....if it's a frozen section, it's been fixed; if it's a P.wax section, it's been fixed in Formalin, fixed in alcohol, subjected to 60C heat.....so, the fact that endog. Peroxidases survive those assaults, indicates that physiologic conditions no longer apply?
>
> Re use of methanol/ethanol....I use IMS. Always have done.
> NB: great excess of substrate can " kill " enzymes ( irreversible substrate inhibition): that is why we add a great excess of H2O2 ( substrate) to "kill" endog. Pxs.
> We then use a stoichiometric quantity of the same reagent ( H2O2) in the presence of DAB/AEC, to visualize sites of AB:Ag interaction.
>
> "For the enzymatic activity of peroxidase it needs an electron-donator"
> The enzymatic activity of peroxidase needs no electron donor, as regards Immuno.....it will "digest" H2O2 into water and oxygen gas.
> However, if you wish to use HRP as a reporter molecule, you must also add a suitable chromogen ( DAB, AEC) that
>
>
> Carl Hobbs
> Histology Manager
> Wolfson CARD
> School of Biomedical Sciences
> Kings College London
> Guys Campus
> SE1 1UL
> Tel: 020 78486813
> Fax: 020 78486816
> 020 78486813
>
>
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> End of Histonet Digest, Vol 104, Issue 15
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