Re: speaking of spleen
you are right. macrophages go around eating up all the excess proteins, so protein
block is a good idea. use normal serum from the host of the secondary antibody and
i also add some serum from the animal species i am staining (ie rat serum for rats,
human serum for humans) i also use serum block before the primary antibody and
then again before the secondary.
Barbara Wright wrote:
> They probably are macrophages - which are not expressing endogenous biotin.
> Endogenous biotin on NBF paraffin sections is usually not a problem unless you
> do HIER. I would suggest you add about 2% serum (of the tissue species) to the
> blocking serum. Or you could use a specific IgG secondary reagent to tag only
> the primary antibody to help rid of this problem. Hope that helps.
> Barb Wright
> Genentech, Inc
> "L. Muffley" wrote:
> > I've been having a terrible time with endogenous biotin in spleen.
> > Yesterday i tried Phyllis Davis' biotin blocking recipie (posted 5/01).
> > The Grocery store concoction of eggs and milk is so wonderfully simple.
> > It did fail however to knock out all non-specific label in my NBF fixed
> > paraffin embedded mouse spleen.
> > The non-specific label i'm getting localizes in cells that look a bit like
> > macrophages. I'm used to looking at skin-not spleen. Does anyone have any
> > suggestions?
> > Thanks!
> > Lara
> > Dermatology Dept
> > University of Washington Seattle, WA
> > email@example.com
> > On Fri, 7 Sep 2001 DDittus787@aol.com wrote:
> > > Almost all tissue has biotin, however :liver , and spleen, seem to be the
> > > worst.
> > > Dana
> > >
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