Brdu incorporation

Jorg Kirberg kirberg at nki.nl
Wed Jan 15 13:09:52 EST 1997


In article <5bgd4i$tus at netnews.upenn.edu>, bhargava at mail2.sas.upenn.edu
(Sangeeta Bhargava) wrote:

> I would really appreciate if someone could give a protocol for the BrdU 
> incorporation. What I exactly want to know is how much and how is the 
> BrdU given in mice. I like to use anti-Brdu in immunohistochemistry to 
> see how the cells are proliferating.
> 
> thanks in advance,
> 
Dear Sangeeta,
maybe you could have a look in one of my papers ? (well, I shoudn't
overestimate myself).
OK, more serious: 
There are two ways used by a couple of groups (Rajewski's lab, von Boehmer's
lab I come from, and many others that I don't have present so
fast), either injecting i.p. or giving BrdU in the drinking water.

For injections: 4 mg BrdU in PBS, inject twice a day (8am and 8pm or something
like that) 200 ul i.p. (adult mice). Keep solution not longer than a week
at 4 C in the dark.

Drinking water: dissolve 0.8 mg (Rajewski's goup) to 1 mg BrdU in the drinking
water; some mouse colonies use acidified H2O which worked in my hands OK.
Wrap the bottle so that it is light protected and renew with fresh prepared
BrdU every three days.

Some mouse strains were better off with the injections I was told, others
prefer the drinking water method. I found both working well and now
use the drinking water one as this way you avoid to stress the mice a lot
which is a desaster with some mouse strains (e.g. DBA/2). However, if
you label for a short time it might be good to start with an
injection because with the drinking water it could take some time
before BrdU is present everywhere.
You should always check whether everything else worked fine, like thymocyte
numbers should stay normal (go down when stressed) and should be labeled
within a short period.

Good luck,
jorg


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Joerg Kirberg                              Tel. 0031 - 20 - 512 19 98
The Netherlands Cancer Institute           FAX  0031 - 20 - 512 20 11
Div. Molecular Genetics (H4)               E_mail kirberg at nki.nl
Plesmanlaan 121
NL - 1066 CX Amsterdam
The Netherlands



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