Re: speaking of spleen

From:rueggp

Barb,
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.
patsy ruegg

Barbara Wright wrote:

> Lara;
>
> 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
> bwright@gene.com
>
> "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
> > muffley@u.washington.edu
> >
> > 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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