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using second antibody with a primary congugated ab


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#1 sara.r

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Posted 12 July 2010 - 02:35 AM

Hi
I already use a primary antibody that is labled with hrp in my elisa but I need to do some flowcytometry too, so can I just use my HRP conjugated ab as primary ab and then add FITC labled secondary antibody? I know the HRP may interfere with interaction of two antibodies but it is hard for me to afford one another unlabled primary antibody.

#2 rkay447

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Posted 12 July 2010 - 10:24 AM

Hi
I already use a primary antibody that is labled with hrp in my elisa but I need to do some flowcytometry too, so can I just use my HRP conjugated ab as primary ab and then add FITC labled secondary antibody? I know the HRP may interfere with interaction of two antibodies but it is hard for me to afford one another unlabled primary antibody.

This should work ok. I've never gone from HRP to a fluorophore secondary but I've used a HRP-conjugated primary and had the signal to weak so I reprobe with the appropriate secondary-HRP and seen a huge increase in signal.




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