It works perfectly fine with 50 ul/well, Lyn. Just be careful to cover the entire surface with the first reagent, since the clean plate might leave some bubbles. Good luck!
You can try 384 well plates and half area well plates which use smaller volumes.
Its not the optimal solution but it works.
50 µl is fine as well. Just shake the plates on a shaker to make sure the the whole buttom of the wells are whetted. The best way at all is the use antibodies with high affinities, only the will decrease the detection limit of your assay so that you can use higher sample dilutions.
I use R&D sandwich to detect mouse IFN gamma, TNF alpha, IL-10 and TGF-beta in serum. I always down-scale the final volume to 50 ul in 96 well flat bottom plates. It works fine. Remember to swirl the plates at every step.
We used to routinely use 50 ul per well for our ELISAs, no problem. Just make sure there are no bubbles - that does cause problems.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA