Optimal developing systems for mouse IFNg ELISpot of HA or IP plates

ZellNet QuestionsCategory: QuestionsOptimal developing systems for mouse IFNg ELISpot of HA or IP plates
David asked 5 years ago

Our lab is trying to decide which plates to use for mouse IFNg ELISpot assays. Either MAHA or MAIPS plates. We would like to know if either plate works better with different developing reagents? For example we are considering Strep-HRP with TMP or Strep-ALP with BCIP/NBT but we are unable to find a resource to indicate if either is mire suited to different plate types.

1 Answers
Sylvia Janetzki Staff answered 5 years ago

Hi David;

Please use MSIP plates. PVDF binds the coating antibody more consistently. It is recommended to prewet the plate with 15 microliters 70% Ethanol, which has to be washed out 3x with PBS (200 microliters), followed by coating. We recommend the Mabtech mouse IFNg kits, they work extremely well. HRP + TMB substrate is an excellent choice. TMB is ta very sensitive substrate. Please use the formulation that is sold by Mabtech as well, since it has to precipitate efficiently to form spots. A good source for answers to your questions is the book: \"Elispot for Rookies (and Experts too)\", published last year by Springer, but available through most other common sources as well.

Good luck! Sylvia