All, ER, clone SP1 is availuable in www.epitomics.com (cat. #4200-1) for $205. Hope this help! Madeleine Huey Supervisor - Pathology (IPOX & Histology) madelein...@elcaminohospital.org
On Fri, Jul 13, 2012 at 7:08 AM, <histonet-requ...@lists.utsouthwestern.edu> wrote: > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-requ...@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-ow...@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > 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 <mw...@wakehealth.edu> > Subject: RE: [Histonet] Epic Beaker module > To: Paula Sicurello <pat...@gmail.com>, HistoNet > <histonet@lists.utsouthwestern.edu> > Message-ID: > > <b2cecb1b6665a4479056478f6de3c4ab0b5e5...@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 > mw...@wakehealth.edu > > > > > > -----Original Message----- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@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@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.vanderl...@amc.uva.nl> > Subject: [Histonet] Re: Methanol in H2O2 explanation > To: "histonet netserver (histonet@lists.utsouthwestern.edu)" > <histonet@lists.utsouthwestern.edu> > Message-ID: > <bda3bc4760afa841926d5fce9d0c16802bd...@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.l...@gmx.at> > Subject: [Histonet] Re: Methanol in H2O2 explanation > To: <histonet@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. > > ________________________________ > > 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. > > > ------------------------------ > > Message: 3 > Date: Thu, 12 Jul 2012 14:48:21 -0400 > From: Brannon Owens <bran...@alliedsearchpartners.com> > Subject: [Histonet] HT/HTL Full Time Opportunities in New York state! > To: "histonet@lists.utsouthwestern.edu" > <histonet@lists.utsouthwestern.edu> > Message-ID: <cc249435.176fe%bran...@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 bran...@alliedsearchpartners.com > <mailto:bran...@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" <ly...@kwbpathology.com> > Subject: [Histonet] ER, Clone SP1 > To: <Histonet@lists.utsouthwestern.edu> > Message-ID: <B422B89C676A4B2F8895D2C4AF3AAE1C@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.ho...@kcl.ac.uk> > Subject: [Histonet] Re: Methanol in H2O2 explanation > To: "histonet@lists.utsouthwestern.edu" > <histonet@lists.utsouthwestern.edu> > Message-ID: > > <11d9615b89c10747b1c985966a63d7ca38626a5...@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 <pat...@gmail.com> > Subject: [Histonet] Beaker module in EPIC > To: HistoNet <histonet@lists.utsouthwestern.edu>, > microsc...@microscopy.com > Message-ID: > <CAPSjddYA=3LUtYM_LA9YmQm1=4St9CjRPCC3Mg8Q=cjoa-a...@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 <jmye...@aol.com> > Subject: [Histonet] p63 and Biocare Medical > To: histonet@lists.utsouthwestern.edu, jmy...@biocare.net > Message-ID: <8cf2e8b2c0332d7-7dc-4...@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" <pru...@ihctech.net> > Subject: RE: [Histonet] ER, Clone SP1 > To: "'Leigh York'" <ly...@kwbpathology.com>, > <Histonet@lists.utsouthwestern.edu> > Message-ID: <6959269F7DA142FCB3EEEC3CFA29B6E2@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 > pru...@ihctech.net > > -----Original Message----- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Leigh York > Sent: Thursday, July 12, 2012 1:06 PM > To: Histonet@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@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_sec...@yahoo.com> > Subject: Re: [Histonet] ER, Clone SP1 > To: Patsy Ruegg <pru...@ihctech.net> > Cc: "<Histonet@lists.utsouthwestern.edu>" > <Histonet@lists.utsouthwestern.edu> > Message-ID: <876c9001-87db-4723-8f71-d99f14d30...@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" <pru...@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 >> pru...@ihctech.net >> >> -----Original Message----- >> From: histonet-boun...@lists.utsouthwestern.edu >> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Leigh York >> Sent: Thursday, July 12, 2012 1:06 PM >> To: Histonet@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@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@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.mor...@ucsfmedctr.org> > Subject: RE: [Histonet] ER, Clone SP1 > To: "Kim Donadio" <one_angel_sec...@yahoo.com>, "Patsy Ruegg" > <pru...@ihctech.net> > Cc: "<Histonet@lists.utsouthwestern.edu>" > <Histonet@lists.utsouthwestern.edu> > Message-ID: > > <8d7c2d242dbd45498006b21122072bf8b5bba...@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-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kim Donadio > Sent: Thursday, July 12, 2012 2:55 PM > To: Patsy Ruegg > Cc: <Histonet@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" <pru...@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 >> pru...@ihctech.net >> >> -----Original Message----- >> From: histonet-boun...@lists.utsouthwestern.edu >> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Leigh >> York >> Sent: Thursday, July 12, 2012 1:06 PM >> To: Histonet@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@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> _______________________________________________ >> 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 > > > > > ------------------------------ > > Message: 11 > Date: Fri, 13 Jul 2012 09:20:25 +1000 > From: "Kim O'Sullivan" <kim.osulli...@monash.edu> > Subject: [Histonet] Gamma delta T cell and CD T cell detection on FFPE > tissue > To: histonet@lists.utsouthwestern.edu > Message-ID: > <CALJ_c9-iXahtW1nTrG2auW8r_U8eVmS=ti-+uu-dmrrfgfw...@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" <pru...@ihctech.net> > Subject: RE: [Histonet] ER, Clone SP1 > To: "'Morken, Timothy'" <timothy.mor...@ucsfmedctr.org>, "'Kim > Donadio'" <one_angel_sec...@yahoo.com> > Cc: Histonet@lists.utsouthwestern.edu > Message-ID: <0227518F256543848B5009F06BDF4EEB@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 > pru...@ihctech.net > > -----Original Message----- > From: Morken, Timothy [mailto:timothy.mor...@ucsfmedctr.org] > Sent: Thursday, July 12, 2012 4:05 PM > To: Kim Donadio; Patsy Ruegg > Cc: <Histonet@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-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kim Donadio > Sent: Thursday, July 12, 2012 2:55 PM > To: Patsy Ruegg > Cc: <Histonet@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" <pru...@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 >> pru...@ihctech.net >> >> -----Original Message----- >> From: histonet-boun...@lists.utsouthwestern.edu >> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Leigh >> York >> Sent: Thursday, July 12, 2012 1:06 PM >> To: Histonet@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@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> _______________________________________________ >> 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 > > > > > > ------------------------------ > > Message: 13 > Date: Fri, 13 Jul 2012 06:45:20 +0000 > From: Jonathan Cremer <jonathan.cre...@med.kuleuven.be> > Subject: RE: [Histonet] ER, Clone SP1 > To: "Morken, Timothy" <timothy.mor...@ucsfmedctr.org>, Kim Donadio > <one_angel_sec...@yahoo.com>, Patsy Ruegg <pru...@ihctech.net> > Cc: "<Histonet@lists.utsouthwestern.edu>" > <Histonet@lists.utsouthwestern.edu> > Message-ID: > > <c35c2709c22fc2458d6a918facc338a30fa30...@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-boun...@lists.utsouthwestern.edu > [histonet-boun...@lists.utsouthwestern.edu] namens Morken, Timothy > [timothy.mor...@ucsfmedctr.org] > Verzonden: vrijdag 13 juli 2012 0:05 > To: Kim Donadio; Patsy Ruegg > Cc: <Histonet@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-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kim Donadio > Sent: Thursday, July 12, 2012 2:55 PM > To: Patsy Ruegg > Cc: <Histonet@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" <pru...@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 >> pru...@ihctech.net >> >> -----Original Message----- >> From: histonet-boun...@lists.utsouthwestern.edu >> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Leigh >> York >> Sent: Thursday, July 12, 2012 1:06 PM >> To: Histonet@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@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> _______________________________________________ >> 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 > > > _______________________________________________ > Histonet mailing list > Histonet@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 <mw...@wakehealth.edu> > Subject: RE: [Histonet] Beaker module in EPIC > To: Paula Sicurello <pat...@gmail.com>, HistoNet > <histonet@lists.utsouthwestern.edu>, "microsc...@microscopy.com" > <microsc...@microscopy.com> > Message-ID: > > <b2cecb1b6665a4479056478f6de3c4ab0b5e5...@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-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Paula > Sicurello > Sent: Thursday, July 12, 2012 4:05 PM > To: HistoNet; microsc...@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@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 15 > Date: Fri, 13 Jul 2012 07:06:34 -0500 > From: "Marsh, Nannette" <n...@stowers.org> > Subject: RE: [Histonet] ER, Clone SP1 > To: "'Jonathan Cremer'" <jonathan.cre...@med.kuleuven.be>, "Morken, > Timothy" <timothy.mor...@ucsfmedctr.org>, Kim Donadio > <one_angel_sec...@yahoo.com>, Patsy Ruegg <pru...@ihctech.net> > Cc: "<Histonet@lists.utsouthwestern.edu>" > <Histonet@lists.utsouthwestern.edu> > Message-ID: > > <2c40e43d1f7a56408c4463fd245dddf9ae46a...@exchmb-02.stowers-institute.org> > > Content-Type: text/plain; charset="us-ascii" > > I love the comparison :-) > > -----Original Message----- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@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@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-boun...@lists.utsouthwestern.edu > [histonet-boun...@lists.utsouthwestern.edu] namens Morken, Timothy > [timothy.mor...@ucsfmedctr.org] > Verzonden: vrijdag 13 juli 2012 0:05 > To: Kim Donadio; Patsy Ruegg > Cc: <Histonet@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-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kim Donadio > Sent: Thursday, July 12, 2012 2:55 PM > To: Patsy Ruegg > Cc: <Histonet@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" <pru...@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 >> pru...@ihctech.net >> >> -----Original Message----- >> From: histonet-boun...@lists.utsouthwestern.edu >> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Leigh >> York >> Sent: Thursday, July 12, 2012 1:06 PM >> To: Histonet@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@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> _______________________________________________ >> 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 > > > _______________________________________________ > 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 > > > > ------------------------------ > > Message: 16 > Date: Fri, 13 Jul 2012 05:19:18 -0700 (PDT) > From: "PATTI NELSON HT\(ASCP\)" <nelsonr...@verizon.net> > Subject: [Histonet] Different Avenues To Become Certified > To: Histonet <Histonet@lists.utsouthwestern.edu> > Message-ID: > <1342181958.20737.yahoomail...@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 > nelsonr...@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-st...@uiowa.edu> > Subject: [Histonet] TRAP staining > To: "histonet@lists.utsouthwestern.edu" > <histonet@lists.utsouthwestern.edu> > Message-ID: > > <17b53f9d0552174bab6f33e0b69bbd9910847...@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_mcma...@urmc.rochester.edu> > Subject: [Histonet] RE: TRAP staining > To: "Sturm, Mary Therese M" <mary-st...@uiowa.edu>, > "histonet@lists.utsouthwestern.edu" > <histonet@lists.utsouthwestern.edu> > Message-ID: > > <ec3287d73321a14799bb96082de57b4753866...@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-boun...@lists.utsouthwestern.edu > [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sturm, Mary Therese > M [mary-st...@uiowa.edu] > Sent: Friday, July 13, 2012 9:18 AM > To: histonet@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@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 <rjbu...@yahoo.com> > Subject: Re: [Histonet] Re: Methanol in H2O2 explanation > To: "C.M. van der Loos" <c.m.vanderl...@amc.uva.nl>, "histonet > netserver \(histonet@lists.utsouthwestern.edu\)" > <histonet@lists.utsouthwestern.edu> > Message-ID: > <1342187715.62239.yahoomail...@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.vanderl...@amc.uva.nl> > To: "histonet netserver (histonet@lists.utsouthwestern.edu)" > <histonet@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.l...@gmx.at> > Subject: [Histonet] Re: Methanol in H2O2 explanation > To: <histonet@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@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 <rjbu...@yahoo.com> > Subject: Re: [Histonet] Re: Methanol in H2O2 explanation > To: "Hobbs, Carl" <carl.ho...@kcl.ac.uk>, > "histonet@lists.utsouthwestern.edu" > <histonet@lists.utsouthwestern.edu> > Message-ID: > <1342188462.30207.yahoomail...@web121406.mail.ne1.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > 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.ho...@kcl.ac.uk> > To: "histonet@lists.utsouthwestern.edu" <histonet@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 > > > _______________________________________________ > 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 > > End of Histonet Digest, Vol 104, Issue 15 > ***************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
