Re: [Histonet] endogenous peroxidases
Make sure you use a fairly fresh bottle of H2O2. (no older than 6 months). The stock bottle degrades after it is opened. I have seen people using an older bottle of H2O2 and the results are sub-optimal. Akemi Akemi Allison BS, HT (ASCP) HTL Director Phoenix Lab Consulting Tele: 408.335.9994 E-Mail: akemiat3...@yahoo.com On Apr 22, 2011, at 10:27 AM, Hobbs, Carl wrote: Gimme 5! Block before or after Ag retrieval. If it's Pwax sections make a 1/10 dilution of 100vols ( ~30%) H2O2 in Dist.water. Block for 15mins. Methanol is not neccessary...imho. Carl Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus London SE1 1UL Tel: 020 78486813 Fax: 020 78486816 ___ 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
RE: [Histonet] endogenous peroxidases
So I have been seeing from this post that people block prior to HIER? I have always done it as the first step after for 10 minutes (I use the DAKO enzyme blocker). Is this wrong? Thanks Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Akemi Allison Sent: Friday, April 22, 2011 12:42 PM To: Hobbs, Carl Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] endogenous peroxidases Make sure you use a fairly fresh bottle of H2O2. (no older than 6 months). The stock bottle degrades after it is opened. I have seen people using an older bottle of H2O2 and the results are sub-optimal. Akemi Akemi Allison BS, HT (ASCP) HTL Director Phoenix Lab Consulting Tele: 408.335.9994 E-Mail: akemiat3...@yahoo.com On Apr 22, 2011, at 10:27 AM, Hobbs, Carl wrote: Gimme 5! Block before or after Ag retrieval. If it's Pwax sections make a 1/10 dilution of 100vols ( ~30%) H2O2 in Dist.water. Block for 15mins. Methanol is not neccessary...imho. Carl Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus London SE1 1UL Tel: 020 78486813 Fax: 020 78486816 ___ 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
RE: [Histonet] endogenous peroxidases
My sequence is: dewax (with dishwasher soap) → wash → HIER → wash → block endogenous peroxidase → go tthe IHC protocol. René J. --- On Fri, 4/22/11, sgoe...@mirnarx.com sgoe...@mirnarx.com wrote: From: sgoe...@mirnarx.com sgoe...@mirnarx.com Subject: RE: [Histonet] endogenous peroxidases To: akemiat3...@yahoo.com, carl.ho...@kcl.ac.uk Cc: histonet@lists.utsouthwestern.edu Date: Friday, April 22, 2011, 1:50 PM So I have been seeing from this post that people block prior to HIER? I have always done it as the first step after for 10 minutes (I use the DAKO enzyme blocker). Is this wrong? Thanks Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Akemi Allison Sent: Friday, April 22, 2011 12:42 PM To: Hobbs, Carl Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] endogenous peroxidases Make sure you use a fairly fresh bottle of H2O2. (no older than 6 months). The stock bottle degrades after it is opened. I have seen people using an older bottle of H2O2 and the results are sub-optimal. Akemi Akemi Allison BS, HT (ASCP) HTL Director Phoenix Lab Consulting Tele: 408.335.9994 E-Mail: akemiat3...@yahoo.com On Apr 22, 2011, at 10:27 AM, Hobbs, Carl wrote: Gimme 5! Block before or after Ag retrieval. If it's Pwax sections make a 1/10 dilution of 100vols ( ~30%) H2O2 in Dist.water. Block for 15mins. Methanol is not neccessary...imho. Carl Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus London SE1 1UL Tel: 020 78486813 Fax: 020 78486816 ___ 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
Re: [Histonet] endogenous peroxidases
The endogenous peroxidase has to be blocked (quenching others call this step) only before the procedure because the idea is to avoid the interference of the peroxidase with it. René J. --- On Thu, 4/21/11, Emily Sours talulahg...@gmail.com wrote: From: Emily Sours talulahg...@gmail.com Subject: [Histonet] endogenous peroxidases To: histonet@lists.utsouthwestern.edu Date: Thursday, April 21, 2011, 4:32 PM Hello I know I can block endogenous peroxidases in paraffin sections before antigen retrieval or after it, but which is better? Does it make a difference? Also, since our H2O2 is 30%, do I need to account for the water in it when I'm diluting it in 100% MeOH? (I was going to use a 0.3% H2O2 in MeOH wash for 30 minutes) I'm not even sure how I would account for it, I just thought it might affect the MeOH since it wouldn't be anhydrous anymore. Emily ps put your hand to the screen for an internet high five!! it's national high five day! A great book should leave you with many experiences, and slightly exhausted. You should live several lives while reading it. -William Styron ___ 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
