I am planning an IHC experiment in mouse brain tissue, but all of the primary antibodies the lab has that I want to use in a co-stain are made in rabbit, so I can't use secondary antibodies to fluorescently label the primary antibodies. The lab has fluorescent pre-conjugation kits they use for cultured cell ICC that I'm thinking of using for my IHC experiment, but I'm not sure how well they would work in tissue in terms of signal amplification, for example. Does anyone have experience with whether this could work, and if so, some considerations in preparing the experiment? Thank you!
Alex Eaker ,
I have tried conjugating primary antibodies with different kits. The brightness of the direct conjugates depends on the expression level of this marker. Usually, I found conjugating to PE gives the brightest signal (using the lightning kit from Abcam) which can produce enough amplification for a low expression marker. For a highly expressed marker, any other color (AF488, AF532, AF555, AF647, etc.) (using thermo kit) should work. Hope this is helpful.
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Atomic, Molecular and Optical Physics
- Atoms