I have treated neutralizing antibody against my protein of interest in cell culture soup. What kind of vehicle should I use for the control cell group?
What is your antibody dissolved in? Look at your product information sheet for your antibody and then make a stock solution based on that. For example, a recent antibody I obtained is stored in: 48% PBS, 50% Glycerol, 1% BSA, is pH 7.3 and contains 0.02% sodium azide as a preservative.
The best would be to use either an isotype control, or another antibody (at the same concentration) to a protein which is not relevant to the process you are monitoring.
Ok. thanks. I am using a goat IgG isotype control, as the neutralizing antibody is raised in goat.
Three ideas added to the excellent suggestions:
(1) Use serum from the uninocculated animal or conditioned media from the hybridoma "mother" culture as negative control.
(2) Spike your culture with the protein of interest to prove biological relevance
(3) Perform ELISA or Western on cell lysates and/or conditioned media before and after (2).
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