Re: NF-KB problems

<< Previous Message | Next Message >>
From:Masayuki Miyagishima <> (by way of histonet)
To:histonet <>
Content-Type:text/plain; charset="us-ascii"

Here is the protocol of NF-kB staining, which ended up with just
interstitial staining( no stain inside cell).

We fixed Tissue with 2X parafolmaldehyde for 2 hours and 30 % sucrose
overnight before freezing in isopentane.  Then 10#027#(I5#027#(Jm frozen
sections were
treated with 5 % normal goat serum for 1 hour at RT, washed with PBS/BSA
and incubated with sc 372 anti-p65 antibody for 1 hour at RT, then
washed and incubated with goat anti rabbit fit C antibody for 1 hour at

Should I do antigen retreival, and prolong the time of incubation?
Masayuki Miyagishima, MD
University of Pittsburgh,
Department of Surgery
412-647-2345, and ask the hospital operator to page 3228, please.

<< Previous Message | Next Message >>