[Histonet] taupathies, LB & amyloid: IHC labeling followed by staining question

[Tyrone Genade]

Hello,

I found the following study, http://www.ncbi.nlm.nih.gov/pubmed/12000724,
where sections were probed with alpha-synuclein antibodies and then
developed with DAB. Then the sections were stained with Congo Red for
amyloid. The afore mentioned article references
http://jhc.sagepub.com/content/10/3/355 for the amyloid protocol. I assume
they, the first article, had used the alkaline Congo Red method. The
results look good and I can use this.

Does anyone have any experience in doing such a double labeling experiment?
Are there any problems that could be anticipated? If I use an alkaline
phosphatase chromogen instead, could I expect problems? I am thinking of
trying the MultiView kit by Enzo Life Sciences:
http://www.enzolifesciences.com/ADI-950-101/multiview-mouse-hrp-mouse-ap-ihc-kit/
and perhaps using some of the more vibrant chromogens:
http://www.enzolifesciences.com/platforms/immunohistochemistry/#An-Unrivaled-Selection-of-High-Definition-Chromogens
. Anyone have any references to papers that spell out a protocol? From what
I can make out, the slides were first developed for IHC and then
transferred to NaCl saturated 80% ethanol and then to the Congo Red where
after the slides were dehydrated and cleared. (I assume, I could just as
well mount in an aqueous mounting medium such as glycerol or
mowiol/n-propylgallate?)

Thanks for the answers to my previous question. Seems 10 micron is a good
middle ground for the demonstration of the various pathologies from serial
and double-stained sections.

Thanks
-- 
Tyrone Genade
Orange City, Iowa
tel: (+1) 712 230 4101
http://tgenade.freeshell.org
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