Re: [Histonet] Re: Maleic acid vs. Tris in in situ buffer

From:"Emily Sours"

I've never heard of using maleic acid for DIG in situ's.  This could be
because we have followed Schaeren-Wiemers and Gerfin-Moser (1993) protocol,
which uses Tris-HCl and makes no mention of maleic acid.
In the papers I've read using DIG in situ's (with chick embryonic tissue,
mind you), there has been no mention of maleic acid.  Where have you seen
this in a protocol?
I suggest "if it ain't broke, don't fix it".  When we've tried to make our
in situ's better by using someone else's protocol, we've *always* gone back
to our original protocol because it's much better.  After ten years of using
it, we know it kicks ass.  Well, at least I do--my PI has doubts, but she
comes around on day three. :)
By the way, is this for whole mount or section in situ's?  And if for
sections, for paraffin or frozen, at what thickness? I'm interested in
knowing in case this helps for paraffin in situ's, since we had beautiful
results and then nothing (though the embryos were fixed in toluene, which
didn't work,  instead of xylene, which worked).  Paraffin in situ's would
rock my world.

yes, paraffin in situ's rock my world.  because i love my job.
Yog-Sothoth knows the gate.
Yog-Sothoth is the gate.
Yog-Sothoth is the key and guardian of the gate. Past, present, future, all
are one in Yog-Sothoth.
He knows where the Old Ones broke through of old, and where They shall break
through again.
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