[Histonet] (no subject)

From:"Orlaith Brennan"




> I'm working on apoptosis in ovine osteocytes. We plan on doing TUNEL staining and also Caspase-3 antibody staining. I was wondering how long from the point the animal dies do you have to freeze the samples before the DNA and proteins will start to deteriorate? Is freezing necessarily the best method of preserving the samples, or should we consider an alternative? Has anyone performed either tunel or caspase 3 staining on undecalcified bone sections?  
>  
> We also plan on looking at other proteins within the bone, inparticular nitric oxide synthase. Again, how long before this protein begins to denature? Does anyone have a protocol for the preservation of bone proteins? 
> 
> Is there a commercial storage solution for proteins (similar to ambions RNALater)? If so would this be effective for a couple of hours to allow the samples be transported?
> 
> Has anyone used a large bone biopsy set?
> 
> Any advice?
> 
> Thank You,
> 
> Orlaith Brennan
> 
> 
> 
> 
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