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From:"Connolly, Brett" <>
To:"" <>, "''" <>
Date:Fri, 23 Apr 1999 09:26:12 -0400


You should avoid acid decalcification. I cannot speak for the preservation
of the specific antigens your are looking at, but I think that chelating the
calcium will increase your odds of success. I have done this with whole
mouse heads for other antigens and for in situ hybridization.

-in a large beaker dissolve 36 g sodium hydroxide in 1L dH2O
-slowly add 100g EDTA, allow to go into solution
-add 12.1 g Trizma base
-pH should be 6.8-7.2

place on a stir plate and keep the solution gently stirring

Wrap the heads in gauze with i.d. # (unless you like to view the heads
swirling around) and suspend into the solution by a piece of suture or the
It takes a while depending on the # of heads (at least a week). Maintain the
pH and check them daily until they are soft enough to cut.
I'd be anxious to here what others suggest.


Brett M. Connolly, Ph.D., HTL
Merck Research Laboratories
Dept. of Human Genetics
P.O. Box 4
Sumneytown Pike & Broad St.
West Point, PA 19486
FAX. 215-652-2075

> ----------
> From:
> Sent: 	Thursday, April 22, 1999 10:37 PM
> To:
> Subject: 	MOUSE NOSES (NALT)
> Looking to  fix and decal whole mouse heads and then hopefully do
> histochemistry staining in the (NALT) associated lymphoid area.....for
> both B & T  cells, germinal center and dendritic cell marker. May use
> mouse antibody B220  mouse CD4.and mouse CD8. Would be tickled eosin
> pink if I had any antigens left after all this processing!!!!! I would
> appreciate any and all help......THANKS     BONNIE G. GREER  ST. JUDE
> CHILDRENS HOSPITAL                            MEMPHIS,TN
> EMAIL>>>>Bonnie,         

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