Re: [Histonet] IHC, decalcification preferences
I, personally, think EDTA SUCKS. a) it takes too long b) its not safe - http://en.wikipedia.org/wiki/EDTA#Toxicity_and_environmental_considerations c) its to old-fashioned. Me, I'm a big proponent of Formic Acid decal. Holla off-list if you want me to hit you up with a protocol, yo. From: Amy Porter port...@msu.edu To: histonet@lists.utsouthwestern.edu; Nicole Collette collet...@mail.llnl.gov Sent: Wednesday, April 1, 2009 12:56:43 PM Subject: Re: [Histonet] IHC, decalcification preferences All the mouse femurs and tibias done in our laboratory are decaled in 14% EDTA and we have a high success rate with our immunohistochemistry and Tunel staining. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Nicole Collette collet...@mail.llnl.gov To: histonet@lists.utsouthwestern.edu Sent: Wednesday, April 01, 2009 12:44 PM Subject: [Histonet] IHC, decalcification preferences Hello, all, I am going to be doing some IHC on adult mouse bone, and since I'm new to all this antigen preservation stuff, was wondering the best way to decalcify? Normally, I use high percentage EDTA (17%) and determine end point by weight loss/weight gain. (up until now have been using this protocol for LacZ stains, works like a charm! ) We are comparing several lines of mice with bone mineral density differences, and endpoint is very phenotype and age-dependent. It takes a long time, but the results are worth the wait. I also have Cal-Rite, which is a formaldehyde/formic acid decalcifier, and obviously works much faster. I am leaning toward the EDTA, since it seems to work well for the LacZ, which is sensitive to lots of other processes, but if there's a reason not to, please let me know. Thanks for all your help and support, I have so far been given great advice from this listserv. Sincerely, Nicole Collette LLNL/UCB ___ 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] IHC, decalcification preferences
Hello, all, I am going to be doing some IHC on adult mouse bone, and since I'm new to all this antigen preservation stuff, was wondering the best way to decalcify? Normally, I use high percentage EDTA (17%) and determine end point by weight loss/weight gain. (up until now have been using this protocol for LacZ stains, works like a charm! ) We are comparing several lines of mice with bone mineral density differences, and endpoint is very phenotype and age-dependent. It takes a long time, but the results are worth the wait. I also have Cal-Rite, which is a formaldehyde/formic acid decalcifier, and obviously works much faster. I am leaning toward the EDTA, since it seems to work well for the LacZ, which is sensitive to lots of other processes, but if there's a reason not to, please let me know. Thanks for all your help and support, I have so far been given great advice from this listserv. Sincerely, Nicole Collette LLNL/UCB ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC, decalcification preferences
All the mouse femurs and tibias done in our laboratory are decaled in 14% EDTA and we have a high success rate with our immunohistochemistry and Tunel staining. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Nicole Collette collet...@mail.llnl.gov To: histonet@lists.utsouthwestern.edu Sent: Wednesday, April 01, 2009 12:44 PM Subject: [Histonet] IHC, decalcification preferences Hello, all, I am going to be doing some IHC on adult mouse bone, and since I'm new to all this antigen preservation stuff, was wondering the best way to decalcify? Normally, I use high percentage EDTA (17%) and determine end point by weight loss/weight gain. (up until now have been using this protocol for LacZ stains, works like a charm! ) We are comparing several lines of mice with bone mineral density differences, and endpoint is very phenotype and age-dependent. It takes a long time, but the results are worth the wait. I also have Cal-Rite, which is a formaldehyde/formic acid decalcifier, and obviously works much faster. I am leaning toward the EDTA, since it seems to work well for the LacZ, which is sensitive to lots of other processes, but if there's a reason not to, please let me know. Thanks for all your help and support, I have so far been given great advice from this listserv. Sincerely, Nicole Collette LLNL/UCB ___ 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
