[Histonet] formic acid treatment and alpha-synuclein staining (Tyrone Genade)

[Dorothy Hu]

>
> I used 88% formic acid in Dwater to pretreatment of mouse tissue slides
> before beta-Amyloid and Anti-Glial Fibrillary Acidic Protein (GFAP)
> antibodies IHC. 10 minutes in formic acid with gental shaking before
> blocking step.


You may also check this paper.

Acta Neuropathol.
<http://www-ncbi-nlm-nih-gov.ezp-prod1.hul.harvard.edu/pubmed/?term=C-terminal+alpha-synuclein+immunoreactivity+in+structures+other+than+Lewy+bodies+in+neurodegenerative+disorders.#>
 2000 Mar;99(3):296-304.
C-terminal alpha-synuclein immunoreactivity in structures other than Lewy
bodies in neurodegenerativedisorders.
Takeda A
<http://www-ncbi-nlm-nih-gov.ezp-prod1.hul.harvard.edu/pubmed/?term=Takeda%20A%5BAuthor%5D&cauthor=true&cauthor_uid=10663973>
1, Hashimoto M
<http://www-ncbi-nlm-nih-gov.ezp-prod1.hul.harvard.edu/pubmed/?term=Hashimoto%20M%5BAuthor%5D&cauthor=true&cauthor_uid=10663973>
, Mallory M
<http://www-ncbi-nlm-nih-gov.ezp-prod1.hul.harvard.edu/pubmed/?term=Mallory%20M%5BAuthor%5D&cauthor=true&cauthor_uid=10663973>
, Sundsumo M
<http://www-ncbi-nlm-nih-gov.ezp-prod1.hul.harvard.edu/pubmed/?term=Sundsumo%20M%5BAuthor%5D&cauthor=true&cauthor_uid=10663973>
, Hansen L
<http://www-ncbi-nlm-nih-gov.ezp-prod1.hul.harvard.edu/pubmed/?term=Hansen%20L%5BAuthor%5D&cauthor=true&cauthor_uid=10663973>
, Masliah E
<http://www-ncbi-nlm-nih-gov.ezp-prod1.hul.harvard.edu/pubmed/?term=Masliah%20E%5BAuthor%5D&cauthor=true&cauthor_uid=10663973>
.

Hope this wil help.

Dorothy Hu
HSDM



> Message: 3
> Date: Fri, 10 Apr 2015 15:00:55 -0500
> From: Tyrone Genade <tgenade <@t> gmail.com>
> Subject: [Histonet] formic acid treatment and alpha-synuclein staining
> To: histonet <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
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> CAEYEE3mA4XHP0_6bRSQZqPuJSoHi15vdNZN+VkBrBB2PFWS4BQ <@t> mail.gmail.com>
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>
> Hello,
>
> I'm do some research on alpha-synucleinopathies and how to stain them. I am
> reading, from the Braak et al papers, that the slides need be treated for
> 10 minutes with formic acid. The method is largely absent. I'm a biochemist
> so I understand the formic acid is denaturing and unfolding the aggregates
> and exposing the epitopes. Would this effect the ability to label other
> epitopes such as plaques and tangles, tyrosine-hydroxylase, FOX2A etc..?
> Does anyone have any experience in double-labeling sections for Lewy Bodies
> and other proteins?
>
> In the same paper an alternative method of a 48 hr incubation on the slide
> is suggested. Details of the staining buffer are absent but I'm hunting
> them down and expect to find something like Triton X-100 in it which will
> again unfold the aggregate (a little).
>
> Does anyone have a reliable protocol to share? I'm now several references
> deep* from my original article and am still trying to find the paper that
> describes the method.
>
> Thanks
> --
> Tyrone Genade
>
> *Back in my Biochemistry days we played a game: Hunting for Bradford.
> Essential, select a paper at random, see if it uses the Bradford Assay and
> find out how many papers you had to read before arriving at the original
> paper by Bradford. My then Prof held the record at some 30+ papers...
>
> Orange City, Iowa
> tel: (+1) 712 230 4101
> http://tgenade.freeshell.org
>
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