We use the Serotec Rat anti Human CD-3, and have used DAKO Retrieval Solution as well as Cell Marque Declere and Trilogy all with good results using the microwave or pressure cooker. The pressure cooker was better though. We used the Vector DAB kit and found that if we used the Nickel it was difficult to see the T Cells, but fairly easy using standard DAB, no frills(using THIS antibody).
Russ Ellis <techmanruss@yahoo.ca>
03/04/03 01:53 PM
To: histonet@pathology.swmed.edu
cc:
Subject: mouse lung CD3 immunostain
Hello everyone
I am developing a method for staining formalin-fixed,
paraffin-embedded mouse lungs with an antibody to CD3.
After an initial review of the archives I was hoping
for some feedback with the following:
1. ANTIBODY - Dako, Serotec, Novocastra, other(?).
I'm sure they all work to some degree, but what are
some of the negative aspects?
2. CITRATE BUFFER - should I be concerned about the
length of microwaving time given that lungs are more
fragile than most other tissues?
3.DAKO ARK - does anyone have any experience with this
"time saving" kit that they could provide some pros
and cons? For me, time isn't a factor if the results
are poor.
I look forward to your useful insights. If anyone
knows of a method that they can point me to as a place
to start, I would be very grateful.
Regards,
Russ Ellis
=====
Russ Ellis, B.Sc., M.L.T.
Research Lab Coordinator
St. Joseph's Hospital
Hamilton, Canada
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