[Histonet] re mouse on mouse
Carl Hobbs
carl.hobbs <@t> KCL.AC.UK
Wed Mar 17 10:38:03 CST 2004
I use Dako mouse anti BrdU( and many other mouse monoclonals) often on pwax
mouse tissue, with no problems; lungs, brain, skin and gut. Std goat anti
mouse biotin secondary with streptavidin- biotin px DAB system( Dako again,
no kits). I personally use BSA as protein additive and ignore serum as a
blocker. IMHO, it's probably the way you try to disclose the BrdU that's
causing the background( obviously, if it's endog peroxidase, too
concentrated primary or biotin... that's different). We've tried many ways
to disclose and many give b'ground. Only clean, specific results we got was
after protease digestion, followed by HCL hydrolysis pretreatments.
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