[Histonet] anti-bromodeoxyuridine staining

Kitchell, Marianne marianne.kitchell <@t> novusint.com
Mon Aug 17 12:25:38 CDT 2009

I am having a problem with my anti-bromodeoxyuridine (BRDU) stain. I
have done this for many years on chicken tissues fixed in Notox. 

Now I have rat tissues (Notox again) and sometimes I get everything
staining (all nuclei) and sometimes I get next to nothing to stain. I
have recently started cutting all my sections (paraffin) at 4 um.
Instead of 5um which may be a partial effect. 

I am pretty sure that my problem is in the 2.0 N Hydrochloric acid (HCl)
step and/or the trypsin step. 

It seems that if I use freshly prepared HCl, I get a better response
than if I have previously prepared and stored my 2.0 N HCl or buy 2.0 N
HCl prediluted. This in turn affects the amount of time required in the
subsequent trypsin step. Every time I get a new bottle of HCl I need to
retitrate the whole stain. Any help here so I am not redoing this into
infinity? I am starting to live the movie Groundhog Day.

I have never been able to perform this stain without an additional
trypsin step even though I have seen it done this way in the literature.

Thank you all in advance.



Marianne Kitchell

Novus International

St. Charles, Mo.


The information contained in this message may be privileged and/or confidential
and protected from disclosure.  If the reader of this message is not the
intended recipient, or an employee or agent responsible for delivering this
message to the intended recipient, please be advised that any reading, review,
forwarding, dissemination, distribution or copying, of this communication or
any attachment(s) are strictly prohibited.  If you have received this e-mail in
error, please so notify the sender immediately.  Also, please delete it and all
attachments from any servers or other hard drives.

More information about the Histonet mailing list