Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

I used to rotinely perform antigen retrieval on frozen tissue sections that
were fixed with paraformaldehyde using 10mM NaCitrate for two short rounds in
the microwave (enough to heat the solution without letting it boil) at the
beginning of the IF procedure.  This seemed to work quite well for IF of
certain molecules.  I'm not sure about the exact mechanism of the reaction but
I think the hot NaCitrate breaks some of the cross-bridges formed during
formaldehyde fixation.

Best of luck!

Mike

>Milton Charlton wrote:
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Dear confocal list members:
>>
>>         In an immunocytochemistry experiment I am having trouble with
>> antibodies for a particular protein that we expect to be present.  We have
>> tried several antibodies for this particular protein using paraformaldehyde
>> or Histochoice fixation with minimal concentration of Triton X or even no
>> detergent.  We also tried incubating the tissue with the primary antibody
at
>> 40C.  Nothing works.    We get good immunoreactivity for other proteins
>> using other antibodies.
>>         There are many references to Antigen retrieval using non
>> physiological pH, chemical treatment or high temperature before application
>> of primary antibody.  Has anyone found these tricks beneficial when applied
>> to non-embedded tissue or cells (no wax)?
>> Thanks for your advice.
>>
>> Milton P. Charlton, Professor
>> Physiology Department
>> Medical Science Building RM.# 3308
>> University of Toronto
>> 1 King's College Circle
>> Toronto, ON, M5S1A8
>> Canada
>> tel:  416-978-6355
>> fax:  416-978-4940
>> [log in to unmask]
>>
>
>
>--
>Edwin D. Moore, Ph.D.
>Department of Physiology
>University of British Columbia
>2146 Health Sciences Mall
>Vancouver, V6T 1Z3, B.C.
>
>phone:  604-822-3423 or 7719
>FAX:  604-822-6048
>email:  [log in to unmask]

>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>
Michael C. Adams
Research Technician for Dr. Clare Waterman-Storer
Laboratory of Cell Motility Studies
Department of Cell Biology
The Scripps Research Institute
10550 North Torrey Pines Road, CB 163
La Jolla, CA 92037

TEL 858.784.9244
FAX 858.784.7521
EMAIL [log in to unmask]
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