Dear Arlind Mara

Speaking from experience the Immulon plates are definitely the best tricky ELISAs. So I would try those. In general plates don't "age" as long as they're kept sterile (most Immulon plates are irradiated as per manufacturer). You can also try the Immulon 4 HBX, which are even more optimized for their adsorption properties compared to the 2HB.

Please also check:

https://us.vwr.com/store/product/4692267/immulon-microtiter-96-well-plates-and-strips-thermo-scientific

Unfortunately the best plates won't save you from the fact that you might want to detect tiny amounts of your cytokine in question, which may be the case as your standard seems to work with the polysorp plates (which in fact are rather hydrophobic).

https://www.thermofisher.com/ca/en/home/life-science/protein-biology/protein-assays-analysis/elisa/elisa-microplates-plasticware/adsorption-immunoassay-plates.html

You might wanna check the clone of your capture antibody and switch out for another.

Good luck with your experiment!

Michael

Usually, for TNFa.. which is easily stimulated in mouse macrophages, I use whatever EIA-RIA high binidng plates that I have on hand (my favorite is still the Costar ones... but that's me). The ELISA manufacturer usually will have a recommendation in their IFU. You can start with that.

Hi Michael Förster Thank you for your suggestion!

I will give the immulon plates a shot.