[Histonet] RE:IHC in mouse lung

[Randolph-Habecker, Julie]

When working with mouse lung and any other mouse tissue it is important to
first block with a Avidin block and a Biotin block. This will bind the
natural avidin/ biotin that can cause background and sticking on the tissue.
In addition to this block you will want to do a serum protein block.  Using
a 15/5% dilution of normal serums.  The 15%, should be from the same species
your secondary was made in (i.e.. if your using Goat anti Mouse as your
secondary-You would block with Goat serum).  And the 5% dilution will be
mouse serum.  These two serums should be mixed together using antibody
diluent.  Block with the serum at least 10 minutes, rinse, then proceed to
apply the antibody that you are using.

I use this technique in my lab on all of our mouse tissue and it works
great! So by taking these steps, it should clear up any background problems
you may be having doing IHC in your mouse lung.   

Lisa   

Lisa McLemore
Lead Histologist
Experimental Histopathology Shared Resources
Fred Hutchinson Cancer Research Center
1100 Fairview Ave. N, G1-300
PO Box 19023
Seattle, WA 98109-1024
Tel: (206) 288-6898
FAX: (206) 288-1345
lmclemore <@t> seattlecca.org 


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