Dear Naira,

You should air dry your sections for about 1 hour. The OCT will wash off in
the wash buffers. Whether you need to do an avidin/biotin block will depend
upon the tissue, but you should try with and without to test this. You may
need to experiment with fixation, look at the datasheet for the antibody and
follow the recommendations. You may need to use paraformaldehyde fixation or
even plp for which you should be able to find the formula in a good
textbook. I use Tris buffered saline (TBS) for washing, others prefer
phosphate buffered saline (PBS). You will find a good protocol in the pack
insert in any of Vector's ABC kits, which are excellent. these pack inserts
also have a trouble shooting guide to help you.

I hope this helps. If you want a copy of my protocol, please contact me.

Margaret

Margaret Blount
Chief Technician
Clinical Biochemistry
University of Cambridge
Addenbrooke's Hospital
Hills Road
Cambridge
CB2 2QR 

-----Original Message-----
From: Margaryan, Naira [mailto:NMargaryan@childrensmemorial.org]
Sent: Tuesday, March 22, 2005 8:19 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] (no subject)


Hello, all,

 

I am going to do Immunostaining of Frozen Sections .

I have some questions:

1. What to do with slides after remove them from --80 o C freezer?

2. How long the slides have to come to room temperature?

3. How the OCT compound has to be dissolved?

4. What is general protocol for frozen sections?

5. Am I have to apply Avidin/biotin?

 

 

My appreciation for any suggestion,

Naira

 

Naira V. Margaryan, Ph.D, D.V.M.

Research Associate II

Children's Memorial Research Center

2300 Children's Plaza, Box 222

Chicago, IL 60614-3394

Tel: 773-880-4000/5-6740

Fax: 773-755-6594

nmargaryan@childrensmemorial.org
 

 

For Express Mail: 

CMRC, Room C.473

2430 N. Halsted Street

Chicago, IL  60614-4314

 



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