Used HBSS ( containing Ca2+ and Mg2+) + 2% FBSS solution for binding antibody. However, all my result seems kind of awkward. Can 2+ ion effect on binding efficiency? or Not?
help me through this!
Thank you
It can, but depends on the antibody and on the target epitope.
Some MoAbs can be "calcium or magnesium dependent" and react with the antigen in presence (or absence of Ca++ or Mg++), but not in the opposite case.
Ca++ or Mg++ can bind to some epitopes (for example vit K dependent proteins) and modify their configuration.
It is very unlikely. Most cell-friendly buffers such as HBSS use a physiological or close-to-physiological concentration of mono- and divalent ions. Under these conditions antibodies should be fine to bind.
Perhaps try DPBS if HBSS gave you an odd result? What was wrong?
It can, but depends on the antibody and on the target epitope.
Some MoAbs can be "calcium or magnesium dependent" and react with the antigen in presence (or absence of Ca++ or Mg++), but not in the opposite case.
Ca++ or Mg++ can bind to some epitopes (for example vit K dependent proteins) and modify their configuration.
Yes, they can have some effect, especially if the epitope bins or is close to a structure binding Ca++.
You can try the same experiment with adding Na2EDTA to your buffer to check the incidence.
Best regards,
Jean
Thank you all for great comments!
I'll keep it up with my experiment.
God bless to all
Best
Min
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