I do X-Gal staining virtually every day, and prefer it to Immuno. The
reasons are that it is a fairly robust protocol, with room for little
errors, or changes in timing, and i can spot a blue cell at a thousand
paces!!!

It is quicker and more immediate than immuno, which is good for a high
pressure situation, but if you have to have immuno then you can always do a
quick screen with x-gal before doing an immuno run (this works best if the
tissue is on slides, where you can x-gal 1 in 5 or 1 in 10, for example)....

There is a question of senstivity, but seeing as i have yet to optimise the
immuno totally to my satisfaction, i use x-gal much more.......there is then
a definate answer for me, instead of a question of background.......

there are so many variations for x-gal recipes, so i wont share mine, as you
will be overwhelmed, but have fun!!!!

emma carter



> -----Original Message-----
> From:	Patterson, Noelle [SMTP:PattersonN@NMRIPO.NMRI.NNMC.NAVY.MIL]
> Sent:	Wednesday, March 15, 2000 7:35 PM
> To:	Histonet
> Subject:	X-gal solution vs. anti-Beta-galactosidase Antibody
> 
> Hello Histonet,
> 
> I have been reading all of the cross talk about Beta galactosidase
> detection
> with interest.  I have recently been tasked to detect beta galactosidase
> in
> murine tissue.  Of course, I haven't done this before (well, I have done
> it
> in bacterial colonies back in my molecular days).  
> 
> I am curious.  Those of you who have to detect this reagent, do you prefer
> the antibody detection, or the enzyme development method for tissues?
> What
> are your reasons for such preference?  Please include all the pro's and
> con's for each method, as your time allows.  
> 
> I am running my first experiment to detect this today (using the X-gal
> solution detection method).  Wish me luck!
> 
> Another question:  Is there a commercial (or otherwise readily available)
> source for beta galactosidase positive control blocks?  If so, would you
> give me the ordering information?
> 
> Thanks for your time!
> 
> Noelle Patterson, M.S.
> NN-TAB
> Bethesda, Md.  20889
> 
> stressed is desserts spelled backwards.  
> 
> 
>