Christoph, If you search out the original reference for the BU-1 clone, you'll find that it indeed does not require extra DNA denaturation for revealing incorporated BrdU. You will also fins an addendum that reveals the discovery of a nuclease that is the actual active mechanism for denaturing the DNA . . . so, it's not so mysterious. I've tried the BU-1 clone in the past, mostly on mouse cells, with little success. I find the most effective method for BrdU detection has been using acid, but recently we combined acid with heat to very effectively reveal BrdU incorporated into several different cell types, both mouse and human. There are several references using DNases to do this . . . I have no experience there. A obvious disadvantage to using acid or heat is the typical loss of surface antigen labeling, the primary reason for developing a DNAse technique. As an alternative, investigate the SBIP technique (strand break induced photolysis, in Xun Li, et.al. Cytometry, Vol. 20, Number 2, June 1995, pp. 172-180). Phoenix Flow Systems has a nice explanation of this, as well as a kit . . see <http://www.phnxflow.com/> MAK. Christoph Goettlinger wrote: > I have a question concerning a MoAb to BrdU, which is designated BU-1. > Right now I am trying to stain for BrdU as well as for different cell > surface markers. So, not all methods are appropiate. Looking for an > resolution, I found an old dicussion about that topic in the archive of > this forum (dated back to 1995). It was said that with the MoAb BU-1 you do > neither need to fix the cells nor do you need to denature the DNA. I was > really happy to find that messages. But following up the discussion was > desillusionating when I tried to find this clone. > I would be very happy if somebody could provide me with the information > where I can get this clone BU-1 from. Any information is more than welcome. > Thank you very much. > > My work address is: Christoph Goettlinger > University of Cologne > Weyertal 121 > Germany > > email: c.goettlinger@uni-koeln.de -- Mark A. KuKuruga, Managing Director University of Michigan Core Flow Cytometry <http://www.cancer.med.umich.edu/flow_cytometry> phone: 734-647-3216 fax: 734-936-7376 kukuru@umich.edu
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