[Histonet] Immunogold

[RCHIOVETTI <@t> aol.com]

In a message dated 12/30/2004 12:22:26 PM US Mountain Standard Time, 
starostm <@t> ors.od.nih.gov writes:
I am looking for protocols for immunogold staining for transmission EM
purposes.  I would like a protocol for DNA and for histone.  We are
evaluating mouse sperm.  I have not been able to find an antibody in the US.
Matthew,

Setting aside the histone labeling issues for a moment, would the osmium 
ammine reaction be suitable for the DNA work?  It provides a quite intense, 
electron-dense reaction product for TEM, and it avoids all of the problems with 
trying to find an anti-DNA antibody, coupling the AB to some form of gold tracer, 
etc.  

There have been a few studies that indicate the labeling of DNA with osmium 
ammine is preferable to using immunogold, and osmium ammine-B is readily 
available from Polysciences (Product is Osmium ammine-B, Cat. No. 48016-91-7).

Just a thought.

Best,

Bob Chiovetti