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. CONFIDENTIALITY NOTE 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet