Anjan, I am more of a fan of steamer or waterbath than pressure cooker, in my= hands at altitude the pressure cooker can be very harsh and make my tissue= s fall off the slide or get chewed up badly. I know the pressure is s= upposed to be controled by the cooker but in my hands boiling seems to be t= aking place, violent boiling and i do not like it.
are you sure the antibodies work on canine tissue? When testing abs, I have four protocols I run 1. no pretreatment 2. HIER with lph citrate buffer 3. HIER with hph buffer, such as edta 4. EIER, I will start with proteinase K for 5min. usually with one of these, if the ab x reacts with canine tissue you w= ould get some signal, if it is weak i would increase the pretreatment times= for that one approach. If i get nothing from any of them and I know = I am using the optomal dilution of the primary ab for the longest incubatio= n time (sometimes i go to over night incubation of primary) then I really w= onder if that ab works on the species I am trying to use it on. Cheers, Patsy -------- Original Message -------- Subject: [Histonet] antigen r= etrieval-pressure cooker-CD44- CD24-canine tissue From: anjan kumar &= lt;drvet_an...@hotmail.com> Date: Sun, May 24, 2009 1:48 pm To: tr= iple immunohistochem <histonet@lists.utsouthwestern.edu> h= ello everyone, I wanted to know the exact stepwise protocol for antigen = retrieval using a pressure cooker. i tried using some protocol but there is= no staining. Kindly tell me the detailed protocol, like when i should plac= e the slides into the pressure cooker and from when i should count the time= . And i am using mouse monoclonals CD44 & CD24 labvision in can= ine tissue, has anyone got experience with these antibodies as citrate buff= er doesnt at all seems to work. plz send me some information on this. Regards, Dr. Anjan Kumar.K.R M.V.Sc Scholar Dept. of = Veterinary Pathology Madras Veterinary College Chennai-7 <= BR>India email: drvet_an...@hotmail.com Phone: +91-9940475801 ___________________________________________________________= ______ Live Search extreme As India feels the heat of poll season, get a= ll the info you need on the MSN News Aggregator [1]http://news.in.msn.com/National/indiael ections2009/aggregator/default.aspx___________________________________ _____= _______ Histonet mailing list histo...@lists.utsouthwestern.edu[2]http://lists.utsouthwestern.edu/ma ilman/listinfo/histonet <= /DIV> References 1. 3D"http://news.i=/ 2. 3D"http://lists.utsouthwestern.edu/mailman/listinfo/histonet"; _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
