[Histonet] M.O.M frozen section background

[Nicholas David Evans]

Hi All,

Does anyone have any tips on reducing background staining when attempting to detect mouse monoclonal antibodies on mouse tissue? We currently use Vector's M.O.M kit  - it works great on PPFE tissue with nice specific staining after pepsin unmasking, but in frozen tissue the background is much higher than the signal.

I routinely use 4% PFA pre-fix, then 3% H2O2 in MeOH for 3 mins, then all the blocks that come with teh kit. I am using several different cytokeratin Abs on 10 um frozen mouse sections.

The only thing that I could think to try was to dehydrate the tissues, rehydrate and try again with the unmasking, but if anyone has a better idea or commonly deals with this problem, please let me know.

Best wishes
Nick