Re: Proliferating cell markers

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From:Robert Schoonhoven <>
To:"Jill McVee (by way of Histonet)" <>
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Cell proliferation studies are a major part of my work.  I have run
these studies on all sorts of animals including humans and, of course, I
have my favorites (antibodies that is).  In my exprience the two best
endogenous markers are PCNA (PC-10) and Ki-67.  I prefer PC-10 as I have
not yet found a crittter with prolferating cells that it would not
stain.  As she is using BrdU I am assuming that she is not working with
humans.  Both PCNA and Ki67 will stain cells in G1 and G2 as well as S
phase but the cells in S phase can be easly identified if one knows what
one is doing.  Not rocket science but it does require some training to
count and get a consistant Labeling Index (LI) between investigators.

BrdU on the other hand is an exogenous marker (a thymidine analogue),
which is incorperated into DNA during S phase.  It is easly counted.  It
can be administered via three routes:

1-Implanted osmotic mini-pump for 3 to 7 days, most invasive but most
accurate gives a total LI over the number of days the pumps are left in,
lots of literature reff.s

2- I.P. injection 1 to 2 hours prior to sac., minimally invasive and
gives an LI about equal to that of PCNA and usefull for studing tissues
that have a natrally high rate of cell proliferation (ie; gut and
stomach), lots of reff.s in the lit.

3-Administration via drinking water, non-invasive, gives an LI between
the pumps and IP injection, our paper is in process, used by us and
origenated at NIEHS, very few reff.s as yet (we're writing!)

Do not use BrdU if you will be looking at DNA adducts as BrdU does cause
them (it IS mutagenic).

"Jill McVee (by way of Histonet)" wrote:
> Hi histonetters,
>                  I have a Research Fellow who is in need of your global
> expertise. She would like to know if anyone has a proliferating cell marker
> anti-body
> which they use routinely  in  IC and is reliable. At the moment she is
> using anti-Brdu in mouse neuronal tissue  after in vivo injection but would
> like to change to a less invasive method of detecting proliferating cells.
> So if anybody out there has replaced their Brdu with another method please
> let me know.
> Jill McVee
> Histologist
> St.Andrews Uni
> Fife. Scotland.

best regards,
Robert Schoonhoven
Laboratory of Molecular Carcinogenesis and Mutagenesis
Dept. of Environmental Sciences and Engineering
University of North Carolina
Chapel Hill, NC 27599
office 919-966-6343
   Lab 919-966-6140
   Fax 919-966-6123 

Don't go around saying the world owes you a living; the world owes you
nothing; it was here first. 
Mark Twain [Samuel Langhornne Clemens] (1835-1910)

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