|From:||"C.M. van der Loos" |
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
NL-1105 AZ Amsterd am
Date: Fri, 10 Feb 2006 15:11:55 +0100 (CET)_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
From:= Guillermo Palao <email@example.com>
Subject: [= Histonet] Mouse tissue staining with monoclonal mouse Abs.
Hel= p with Vector Mouse on Mouse kit.
To: Histonet <histone firstname.lastname@example.org
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.
Thanks in advance,
Gui= llermo Palao, MD. Ph.D.
Laboratorio de Reumatología
Centro de Investigación
Hospital 12 de Octubre
Avda de Córdoba s/n