But the heat also damage the fluorescnece...
u need specific amplication kit anyway.
2009-05-17
TF
发件人： Gudrun Lang
发送时间： 2009-05-17 02:00:18
收件人： ti...@foxmail.com
抄送： histonet@lists.utsouthwestern.edu
主题： AW: [Histonet] Chemicals that inactivate the primary antibody
For doublest
For doublestaining the primary undergoes denaturation through a second
HIER-step. The second secondary ab doesn't bind to the first primary.
Therefore the binding sites must have been destroyed by heat in
retrieval-buffer.
Gudrun
-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsou
I did try the frozen section from fresh tissue...and fix them in cold acetone
for 10 min, transfer to cold PBS for 30 min, then wash in PBS at RT, begin IHC.
But the tissue quality is really to bad. You always saw pieces of
tissues floating, fall in to small pieces and gone when you wash
Hi, but I dont want to destory everything
I am trying to see if I can prevent the binding of another 2nd antibody to the
primary antibody for the first antigen!
There fore you can perform double staining from same sepecies-derived primary
antibodies!
2009-05-17
TF
发件人： Rene J Buesa
The NSH IHC Resource Group is also a good source for info on IHC, NSH
members can join the group online at www.ihcrg.org
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
email pru...@ihctech.net
websi
No you need to thaw/fix in formalin or alcoholic fixative in order to
process and embed in paraffin.
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
email pru...@ihctech.net
website www.ihctech.
U would be better off freezing the brain unfixed than trying to fix in alcohol
and freeze it, alcohol fixation is not recommended for freezing tissue. Just
freeze slices of the brain or a whole rat or mouse brain and then make sections
and fix the sections in alcohol/acetone mixture. Sectionin
Try bleach and you will destroy everything!
René J.
--- On Sat, 5/16/09, TF wrote:
From: TF
Subject: [Histonet] Chemicals that inactivate the primary antibody
To: "Histonet@lists.utsouthwestern.edu"
Date: Saturday, May 16, 2009, 12:48 PM
Hi all, just wonder what kind of treatments/chemicals
Hi all, just wonder what kind of treatments/chemicals can complete block the
binding of 2nd antibody to binded primary antibody on antigen?
I tried HCl , but does not damage all the primaryantibody binding sites - i can
still see staining pattern finally.
2009-05-17
TF
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