I'm sorry Joyce, I sometimes forget.  RxM and MxR stand for rabbit
anti-mouse and mouse anti-rabbit, respectively.

Linda A. Sebree, HT
University of Wisconsin Hospital & Clinics
Immunohistochemistry/In Situ Hybridization Laboratory
D4/218-2472
600 Highland Avenue
Madison, WI  53792-2472


(608)265-6596
FAX: (608)263-1568

> -----Original Message-----
> From:	Joyce Kotzuk [SMTP:JKotzuk@salud.unm.edu]
> Sent:	Thursday, November 11, 1999 11:03 AM
> To:	Yuhui_Xu@hms.harvard.edu; la.sebree@hosp.wisc.edu;
> histonet@pathology.swmed.edu
> Subject:	RE: signal amplification methods
>
> Could someone please explain this to me. What is RxM IgG? Thanks, Joyce
>
> >>> "Sebree Linda A." <la.sebree@hosp.wisc.edu> 11/10/99 11:31AM >>>
> Hi Yuhui,
>
> We use Ventana automated instruments so we have the option of using their
> Amplification Kit.  This is comprised of RxM IgG followed by MxR IgG. This
> is applied after the primary and before the secondary.  In essence, you
> are
> amplifying the binding sites of the primary antibody.  If this is not an
> option, you can achieve similar results by making your own reagents.  We
> use
> RxM IgG (Pierce, cat. #31190) at a 1:200 dilution in between the primary
> and
> secondary antibodies (you could probably follow this with a MxR IgG
> although
> we haven't tried this).  This is sometimes slightly dirtier than the
> Ventana
> Amplification Kit but its a heck of a lot cheaper!  And one could
> certainly
> play with the dilutions and incubation times to clean it up.
> Unfortunately,
> you are amplifying any endogenous peroxidase and/or biotin not just the
> primary antibody sites, so you should make sure you have a nice clean
> stain
> before you amplify. Obviously these methods are for use with monoclonal
> antibodies although we have tried both our home brew amplification and
> Ventana's with polyclonal antibodies and sometimes they do intensify the
> reaction!
>
> Hope this is of some use,
>
> Linda A. Sebree, HT
> University of Wisconsin Hospital & Clinics
> Immunohistochemistry/In Situ Hybridization Laboratory
> D4/218-2472
> 600 Highland Avenue
> Madison, WI  53792-2472
>
>
> (608)265-6596
> FAX: (608)263-1568
>
> > -----Original Message-----
> > From:	Yuhui_Xu [SMTP:Yuhui_Xu@hms.harvard.edu]
> > Sent:	Wednesday, November 10, 1999 7:56 AM
> > To:	histonet@pathology.swmed.edu
> > Subject:	signal amplification methods
> >
> > Dear All:
> >
> > I understand that currently there have been methods to amplify
> > immunohistochemical signals, e.g.,TSA by DuPont. I would appreciate
> > information
> > about other methods in this direction.
> >
> > Thanks in advance.
> > Yuhui
> >
>