Re: [Histonet] Antigen retrieval

From:"Victoria Baker"

Rene and Kim

It's not a routine procedure, but what Kim hasn't said is why this is
needed.  Yes in certain instances - histochemically speaking - some
incubation's need to be done overnight but not at quite that high a
temp.  Immunohistochemically - I've done it but it took time to work
out the best method to achieve

When we've had to do these it was done in a waterbath for no more than
12 hours and the bath covered loosely either by a tent cover (it comes
with the waterbath)  or aluminum foil to hold the humidity in.  The
amount of buffer required for the slides will be pretty hefty, but
it's more important to make sure you have a waterbath that can
actually maintain the temperature and surrounding moisture so that the
buffer won't evaporate.  This is going to require some testing during
the day to see how best to make this work.  Kim if you are looking to
do a large volume of slides make sure you use a pyrex dish that fits
into the waterbath or one that can be supported in a bath - sort of
like using a double boiler when you're melting chocolate (sorry for
the less than scientific description)- so that you don't burn out the
heating element.  I also put in blank slides to best mimic the volume
of work I'd be doing.  The bath was set up first thing in the morning
and both the temp and water levels would be monitored during the day
this way we could best gauge and write up our protocol.  I know that
there is literature on doing this procedure by Dr. Elias, but I don't
have it in front of me.

I hope this helps, if there is anything else I can do please let me know.

Vikki Baker

On 4/12/08, Rene J Buesa  wrote:
> Never heard of such a procedure. On the other hand, unless you are dealing with a HUGE amount of buffer, it will evaporate during such a prolonged period of time.
>  Check your reference, I think something is not right.
>  René J.
> Kim O'Sullivan  wrote:
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