Deirdremast 2015-12-08 19:30:44
I am a research scientist, currently trying to optimise an ELISA assay
for serum interferon alpha. I am using a mouse derived
monoclonal(primary) antibody with an unlabelled rabbit
polyclonal(secondary) antibody.In order to produce an adequate
enzymatic change, I am using a biotinylated goat anti rabbit
anti-immunoglobulin followed by extravidin-AP and pNPP. However, at
lower concentrations I am getting a huge amount of background.
Does anyone have any ideas where and what to introduce as a second
blocking step to reduce the amount of backgrond.