rat and mouse immunostaining/compiling info

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
Date:Thu, 24 Jun 1999 10:25:20 -0600

Good Morning Histonetters,

I am in the mood to put together a great deal of information on immunostaining
mouse and rat tissue, primarily cell (CD) markers and once compiled, 
put it out on Histonet.  Any takers???  

I know that most markers will work with frozen sections but many people
DO NOT or ARE NOT interested in working with snap frozen tissues since they
already have tissues fixed/paraffin embedded and utilized for other staining
protocol, multiparametered labs.  

The thrust will be NBF or other fixation with paraffin sectioning.

I will need and please be very specific/detailed:


processing schedule in paraffin:

Antigen retrieval and/or enzyme digestion used:  

Buffers used in protocol/detergents used.

peroxidase block used:  concentration/solvent in blocker

normal serum or other blocking

avidin biotin block

antibody, diluent used, time, temp of incubation

secondary, diluent used, time, temp of incubation



Special "tweaking" if applicable

Am I asking for too much?  These are frequently asked questions (FAQ)
and may be a great help to many getting started.  

Do you want to include frozen section work, now or separately, this will be 
indicated if you have had trouble with NBF fixed tissues.

I will not include antibody vendors, but will give specifics on antibodies
used, example:  secondary antibody, goat xRat, F(ab')2 absorbed to mouse,
human, etc., you can shop on your own or ask more questions.

I know Jamie Erickson will arise to the occasion as she and many others
answer these questions frequently.  The need is there as more of us are
involved with these research animals. In all fairness, would like to
acknowledge people who contribute.  

IS there anything else you would like included??  

Gayle Callis

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