Using ECF for protein detection? - ECF instead of ECL? (May/27/2006 )
in the ECF system instead of luminescence emission, like in ECL, there is Fluorescence emission.
The secondary antibodies are linked to Alkaline Phosphatase (AP).
The AP catalyses the conversion of ECF substrate to a highly fluorescent product which fluoresces strongly at 540-560nm.
So far , I am not very happy with this system because is not as sensitive as the ECL system used with films. The signal I obtain is significantly lower.
What I would like to know is:which laser or filter should I use?
Anyone using ECF?