Dear  Guillermo,
  
Perhaps you should
   try  a biotin-free detection system instead of t= he Vector MOM kit.
   In  my  hands  endogenous  biotin  cannot be effectively = quenched in
   kidney tissue. Molecular Probes/Invitrogen has a Zenon kit = that is
   in  vitro labeling your mouse primary antibody with either enzymes= or
   fluorochromes via a labeled Fab anti-mouse reagent. Then c   ontinue either with fluorescence microscopy or anti-FITC for enzymatic
   de=  tection.
 
Hope this helps.
 
Chris
   van  der  Loos,  PhD
Dept.  of  Pathology
Acade= mic
   Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterd   am
The       Netherlands
      
 
   
Date:    Fri,    10    Feb    2006    15:11:55    +0100
   (CET)
From:= Guillermo Palao
   <gpbnas@yahoo.es>
Subject: [= Histonet]
   Mouse  tissue staining with monoclonal mouse Abs.
Hel= p with
   Vector  Mouse  on  Mouse kit.
To: Histonet <histone   t@lists.utsouthwestern.edu

Dear
   all,
&nbs=   p;  
I  am  working
   with mouse frozen sections o= f different autoimmune disease models,
   implying an important mouse IgG = deposition in studied tissues (mouse
   kidney).  For  this  reason  we bought= Vector Mouse on Mouse kit to
   avoid  detection  of  deposited  IgG  by  seconda= ry antibodies while
   maintaining   sensitivity   for   mouse   primary  antibody.=  After
   increasing  concentration  of  blocking reagent as well as decreasing   concentration  of  biotynilated  anti-mouse  IgG,  there  still is a
   "back=  ground"  of  positive  staining  of deposited IgG in renal
   glomeruli togeth= er with specific staining of antigens of interest.
   I  would  really  appre=  ciate  any  tips  or  ideas  you might think
   helpful. 
 &n= bsp; 
  Thanks in
   advance,


Gui=     llermo    Palao,    MD.
   Ph.D.
Laboratorio   de  ReumatologíaCentro  de
   Investigación
Hospital    12   de   Octubre
Avda   de   Córdoba s/n
Madrid
   28041
Spain

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