[Histonet] BrdU on fixed frozen section

Johnson, Teri TJJ <@t> Stowers-Institute.org
Mon Jan 23 12:58:50 CST 2006


Eva,

Are you denaturing them in HCl prior to BrdU antibody? This antibody
requires denaturing by HCl or microwaving in order to recognize the BrdU
epitope. DNA in cultured cells and frozen sections should be denatured
in 2-4M HCl treatment, rinsed, and then incubated in antibody. 

If you are currently doing this and still get no signal, send me your
protocol (including dilutions) and we can go from there. (Direct
detection vs. indirect detection) How long are your samples fixed prior
to freezing?

Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64133



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