[Histonet] Double stain IHC question

[Steven Weston]

This is probably most easily done using immunofluorescence since it works well on frozen tissue. You could use a streptavidin linked FITC tag for the human biotinylated Ab, and then use a red tagged anti IgG directed against the species of the other antibody. (hoescht reagent will stain the nuclei)

If however you want to use immunohistochemistry you can use a strepatvidin linked alkaline phosphatase with BCIP/NBT (blue chomogen) for the biotinylated Ab and a polymer based peroxidase secondary for the other antibody and visualise using DAB. For nuclear staining you can use nuclear fast red. I have used this method recently to stain cytospots successfully for similar macrophage markers (inos and cd68).

Hope this helps.

regards

steve weston
lab manager
Breathe-Well CRE
UTAS-SOM


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