[Histonet] beta gal antibody

[Mesru T]

Hi James,

For IHC: I fix the tissue after grossing in 4% PFA ( 10% NBF should work
fine as well), then proecess for paraffin and embed. I have never tried
this marker with gluteraldehyde fixation. Usually gluteraldehyde is not
suggested for IHC.

Mesru
On Fri, Jan 3, 2014 at 2:26 PM, James Watson <JWatson <@t> gnf.org> wrote:

> Do you still fix with the 4%pfa + 0.2% glutaraldehyde or can the tissues
> be fixed in formalin for paraffin sections?
>
> James Watson HT  ASCP
> GNF  Genomics Institute of the Novartis Research Foundation
> Tel    858-332-4647
> Fax   858-812-1915
> jwatson <@t> gnf.org
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:
> histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Mesru T
> Sent: Friday, January 03, 2014 10:59 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] beta gal antibody
>
> Hi Elizabeth,
>
> I always read and benefit from the messages here. Now I so happy that I
> think I might be of some help to you or also others.
>
> When I do enzymehistochemistry to detect the beta galactosidase enzimatic
> activity using x-gal as a substrate, I mostly use fresh frozen tissues and
> fix the frozen sections in 4% PFA + 0.2% gluteraldehyde in PBS for 5min.
> When I do IHC I prefer to use paraffin sections (the morphology is
> superior and I find it easier to cut paraffin than frozne).
> Frozen sections wll work for IHC too. The enzyme does not need to be
> active for antibody to bind during IHC satining.
>
> I have tried many antibodies until I have found the one that works for me
> on FFPE mouse tissues: Beta-galactosidase  from eBioscience cat#14-6773
> lot#E05196-1631  I use it at 1ug/ml concentration.
>
> I hope this helps.
>
> Regards,
> Mesruh Turkekul
> mskcc.org
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