Hi guys,
I am going to conduct an affinity purification experiment using a synthetic peptide coupled to an agarose matrix to pick up the specific Abs from the pAb fro my rabbit serum, my question is; shall I dilute the serum sample before loading into the column, and what kind of diluent I should use ?
NB. I am using a 1 ml cartridge.
Dear Haitham
Please see the attached article for further describtion
for viscosity reasons, in order for it to flow well, you could dilute in something like PBS, but I don't think you need to do that. It wont hurt to dilute and could help non specific sticking to the column. Since it is an affinity column, it will re-concentrate on the column if you dilute it out. I would look at the literature provided by the Thermoscientific website for purification of antibodies.
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