Methods and Techniques Discussion > Immunofluorescence (IF)

How two use two primary raised in same species in IFC

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rima:
Is there any way to use two primary Antibodies raised in same species.
My both primary are raised in Rabbit. Your expert help will be appreciated.

TMazurek:
I have the same issue right now as well. First, I tried looking for another primary I could swap out, but unfortunately I had no luck. My next thought is to possibly incubate the primary antibodies with secondaries before IFC so that I create primaries with secondaries already attached. I could also purchase this already made (my target primary with an Alexa Fluor conjugate attached), but I have no previous experience with this and don't know how well it would work when primaries will have matching hosts. I would love to hear if anyone else has tried this or what other techs did to get around this.

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