1 reply to this topic

[BioMiha]

Hi all,

I am curious if anyone has similar experience. I am observing a lot of anti-avidin (or more specificaly NeutrAvidin from Pierce) antibodies in normal human sera. I am trying to devise an assay that uses a biotinylated antigen but I can't seem to loose the background signal. I don't want to dilute my target sera too much so as to be able to observe low affinity specific Abs as well.
Thanks for the feedback.
Miha

[antibodymania]

I used to run a control sample with D-Biotin from Thermo and measure the background, then add my test sample. Any additional signal should be from the target biotin - D-biotin is supposed to get kicked off when biotin is present.
Alternatively, you could run 2 samples in parallel: 1 with d-biotin and 1 without and just subtract the background.