Effect of DTT on the affinity of Protein A/G and immune complex - Effect of DTT on the affinity of Protein A/G and immune complex (Nov/29/2010 )

Dear everyone,

We are performing a proteomic approach on the ICs, my colleagues suggest an "on beads" treatment of DTT and Iodoacetamidd for immune complex before elution step.

Do you have any idea how these DTT and Iodoacetamidd affect on the binding of immune complex and protein A/G?

Thanks

Depends on what you will be doing with the eluate. Protein A/G is extremely stable so you will not release it (at least not considerably) from the beads.

Correct me if I'm wrong, but will the DTT not break the disulfide bridges that keep the heavy chain and light chain of your antibody together? Maybe it depends on the concentration, but I think you should be careful in using a reducing agent before the elution step.