EDC/NHS problem

Hi,
We used gold-covered glass chip and we incubated in MUA-11 in overnight. While we activated the chip surface with EDC/NHS, we saw an unusual binding problem while incubating. We want to get your opinion about our graphs and binding. what do guys think about our Bsa binding and anti bsa binding graph ? What are you suggesting us to do? Thank you.

Hi,
I am not familiar with the scale of the sensorgram but when I compare the relative heights I think that the binding of the BSA (68 kDa) compared to the activation (NHS/EDC < 1 kDa) is very low. In addition the injection of anti-BSA (150 kDa, 1 µg/ml = ~7 nM) is not giving a binding curve which also may indicate insufficient BSA is bound or the antibody concentration it too low.
So try to bind more BSA and/or raise the anti-BSA antibody concentration.
Kind regards
Arnoud