[Histonet] X-gal staining

[Amos Brooks]

>
>
> Hi,
     I'd like to concur with Carl Hobbs. With formalin fixed, paraffin
embedded tissue, beta Galactosidase is definitely the way to go. It is just
an antibody so you would do it like any other IHC.

Amos


Hi
> No replies so far so.....my pennyworth.
> Imho ...no
> beta Gal enzyme is inactivated by std Pwax processing.
> So, when you apply the X-gal substrate and chromogen solution to Pwax
> sections,  it will not give you a positive blue final reaction product
> If you want to detect beta Gal in FFPWS and visualise with DAB,  you will
> have to use an anti beta Gal antibody.
> Finding an anti beta Gal enzyme antibody, that works in FFPWS +/- AR is
> not so easy.
>
> Good luck
>
> Carl
>
> Carl Hobbs FIBMS
> Histology and Imaging Manager
> Wolfson CARD
> Guys Campus, London Bridge?
> Kings College London
> London
> SE1 1UL
> ?
>
> 020 7848 6813
>
>
> Subject: [Histonet] X-gal staining
>
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> Hello histonetters,
>
>
> I have request from a scientist to perform X-gal staining on the formalin
> fixed paraffin-embedded kidney.
>
> I was wondering if there is any possibility to do this staining on FFPE
> tissue as well as if there any possibility to perform a peroxidase DAB
> staining for the X-gal? If so, may I ask you for a detailed protocol to
> perform this staining.
>
> Any input and explanation why it is impossible is appreciated.
>
>
>
> Thanks in advance,
>
> Naira
>

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