[Histonet] antigen retrieval-pressure cooker-CD44- CD24-canine tissue

[pruegg <@t> ihctech.net]

   Anjan,

   I  am  more  of a fan of steamer or waterbath than pressure cooker, in
   my   my  tissue   pressure  is  s   boiling  seems to be t   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   pretreatment  times   of them and I know    for  the  longest  incubatio   incubation  of primary) then I really w   species I am trying to use it on.



   Cheers,

   Patsy



   -------- Original Message --------
   Subject:   [Histonet]   antigen   r   CD24-canine
   tissue
   From: anjan kumar &   Date: Sun, May 24, 2009 1:48 pm
   To: tr   h   I  wanted  to know the exact stepwise protocol for antigen    using  a pressure cooker. i tried using some protocol but there is   staining.  Kindly  tell  me  the detailed protocol, like when i should
   plac   count the time   And  i  am  using  mouse monoclonals CD44 & CD24 labvision in can   tissue,  has  anyone  got  experience with these antibodies as citrate
   buff   plz send me some information on this.
   Regards,
   Dr. Anjan Kumar.K.R
   M.V.Sc Scholar
   Dept. of    Madras Veterinary College
   Chennai-7
   <   email: drvet_anjan <@t> hotmail.com
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