[Histonet] Primary works with ABC but not IF

Patten, Nicole (NIH/NIAAA) [F] pattennj <@t> mail.nih.gov
Wed Dec 10 09:09:45 CST 2008


I am in a situation where my primary antibody works using the ABC
method, but I do not see staining with immunofluorescence. My IF
protocol works well with other antibodies so I do not know why it's not
working with this particular antibody. Does anybody have any

I have considered using an avidin-Alexa Fluor conjugate but I am not
sure if this is even possible (I am new to IHC) or how I would go about
it. Would I need to first biotinylate my primary?

Does anyone have any advice/protocols? 

Thanks! Any help would be greatly appreciated.

Nicole Patten
Post-Baccalaureate Fellow/IRTA
NIAAA/National Institutes of Health

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