floating section antigen processing

[Feinstein, Timothy]

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Hello all,

This is only semi-relevant but a friend needs a good protocol for antigen
retrieval involving floating sections of previously frozen brain.  She has
seen several published protocols but the first one she tried (Jiao et al.,
J Neuroscience Methods 93: p149) produced unsatisfactory results and she
needs to keep trial and error to a minimum if possible.  If you have any
favorite protocols or can point to a reference you find especially helpful
please let me know off-line so as not to tie up the listserv.

Many thanks,


TF

Timothy Feinstein, Ph.D. | Confocal Manager
333 Bostwick Ave., N.E., Grand Rapids, Michigan 49503
Phone: [phone] | Email: [hidden email]

[Linda Barthel]

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Tim,
What is she trying to localize?  A 2N HCl pretreatment can work on some
cases.  It isn't as reliable as hot sodium citrate but it could easily be
applied to sections.  Brain tissue is very heavy in fatty tissues so adding
a mix of triton, tween and DMSO to the ICC reagents may help

Linda Barthel
University of Michigan
Dept. Molecular, Cellular and Devlp. Bio.


On Monday, August 18, 2014, Feinstein, Timothy <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hello all,
>
> This is only semi-relevant but a friend needs a good protocol for antigen
> retrieval involving floating sections of previously frozen brain.  She has
> seen several published protocols but the first one she tried (Jiao et al.,
> J Neuroscience Methods 93: p149) produced unsatisfactory results and she
> needs to keep trial and error to a minimum if possible.  If you have any
> favorite protocols or can point to a reference you find especially helpful
> please let me know off-line so as not to tie up the listserv.
>
> Many thanks,
>
>
> TF
>
> Timothy Feinstein, Ph.D. | Confocal Manager
> 333 Bostwick Ave., N.E., Grand Rapids, Michigan 49503
> Phone: [phone] | Email: [hidden email]
>

--
Linda Barthel, M.S.
*Research Laboratory Specialist Lead*

*Department of Molecular, Cellular, and Developmental Biology*
                      3010 Natural Sciences Building (Kraus)
                                    830 N. University
                             Ann Arbor, MI  48109-1048
lab: [phone]
fax: [phone]
http://www-personal.umich.edu/~praymond/

      *  Microscopy & Image-analysis Laboratory-North *
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office: [phone]
fax:    [phone]
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[Michael Delannoy]

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Timothy,
Try this reference:
Microscoscopy Immunohistochemsitry, and Antigen Retrieval Methods
For Light and Electron Microscopy

By: M.A.Hayat
2002  ISBN 0-306-46770-4
Kluwer Academic/Plenum Publishers N.Y.

Good luck,
Michael Delannoy

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Feinstein, Timothy
Sent: Monday, August 18, 2014 7:45 PM
To: [hidden email]
Subject: floating section antigen processing

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Hello all,

This is only semi-relevant but a friend needs a good protocol for antigen
retrieval involving floating sections of previously frozen brain.  She has
seen several published protocols but the first one she tried (Jiao et al., J
Neuroscience Methods 93: p149) produced unsatisfactory results and she needs
to keep trial and error to a minimum if possible.  If you have any favorite
protocols or can point to a reference you find especially helpful please let
me know off-line so as not to tie up the listserv.

Many thanks,


TF

Timothy Feinstein, Ph.D. | Confocal Manager
333 Bostwick Ave., N.E., Grand Rapids, Michigan 49503
Phone: [phone] | Email: [hidden email]