I am having trouble getting consistente results with MAIPS4510 ELISPOT plates (Merck, Millipore). We have used them before with no trouble and using the same protocol. Or positive controls (PHA) does show spots but less then expected.
- I tried fresh and frozen PBMCs, also from healthy donors.
- I checked and reordered all media, reagens and antibodies to make sure they are not out of date or stored inappropriately at some point.
- I use the pre-wetting as per user manuel, and washing without Tween.
I have the feeling more people use MSIPS4510 from Merck, but according to the manufacturer the only difference is the underdrain to make it accesible to robots; no difference in filter.
Does anyone have an idea of what I could still try to improve my ELISPOT?
Thank you!
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