Re: bone and IHC


i use zinc formalin fixation and 5% formic acid (Immunocal from Decal Chemicals) decalcification and then routine paraffin processing for IHC on bone samples.  the zinc formalin will elliminate most need for HIER which is really hard on bone sections.  formic acid decal has been shown to do the job without adversely affecting the antigen sites, it is much faster than EDTA (which can affect proteoglycans) but slower than HCL or Nitric decalcifiers which can be hard on antigen sites.  the sections tend to come off the slide easily unless you carefully drain them before heating and of course good quality silane coated plus slides are required.   if you can avoid HIER it is suggested.  i prefer to use enzyme digestion EIER with bone IHC, I find it easier to control.  my preferred enzymes are proteinase k ready made from DAKO and pepsin ready made from Research Genetics.
patsy ruegg wrote:

I was wondering how people out there process bone samples that will be used for immunohistochemistry - specifically rodent specimens. Any insights would be most appreciated. And please try to cc me directly as well as the Histonet. Thanks!Andrea

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