Re: [Histonet] Immunogold
In a message dated 12/30/2004 12:22:26 PM US Mountain Standard Time,
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.
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,
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.
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