Hi all

I am doing an experiment whereby I extract IgG from serum using an affinity collumn assay (Protein A- columns: NAb Spin Columns, Thermo Scientific) , and I want to expose cultured endothelial cells to the negative fraction of serum (IgG-depleted serum).

However, per protocol the serum has to be diluted (1:4) before passing through the Protein A- columns (NAb Spin Columns, Thermo Scientific), and the effect I am looking for is lost when cells are exposed to the diluted serum.

So two choices:

a) I try to pass undiluted serum through the columns

b) I try to "re-concentrate" the negative fraction through dialysis with a concentrating gradient (such as Spectra/Gel Absorbent with FLoat-a-Lyzer G2, Spectrum Labs).

Do you have any experience with any one of these alternatives?

Best,

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