[Histonet] Lac Z staining

[Stefano Mantero]

Hello Julie,
Unfotunately the beta Galactosidase are inactivated
from heat. If you want to use the paraffin you have
two possibilities:
1 reveal the beta galactosidase with an antibodies
(Chemicon works well)
2 use a very low melting point paraffin (42-44 C), in
this case you preserve the activity of the enzyme but
the section is not simple to obtain, it is necessary a
cryostat setted at +4 C

Good luck
Stefano


Stefano Mantero
Dulbecco Telethon Institute
Via fratelli Cervi, 93
Segrate (MI)
Italy

  --- Juile Gorenstein <j_gorenstein <@t> yahoo.com> ha
scritto: 
> Hello everyone,
> Thank you to those who has responded to my embryo
> processing e-mail!
> I was wondering if anyone has a protocol for lac Z
> staining on paraffin sections.  I have been staining
> frozen sections, but have been having a little bit
> of trouble keeping the tissue attached to the slides
> (I use Superfrost Plus slides:  I allow the tissue
> to dry for about an hr after I attach the section to
> the slide, store them at -80oC and then dry them
> again before staining -- I stain by hand, it's not
> an instrument issue).  Please let me know if a
> protocol for paraffin sections exists.
>  
> Thank you in advance,
> Julie Gorenstein
> Novartis, Cambridge, MA
> 
> 		
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