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The percentage of binding was determined by measuring the radioactivity bound to the rhCEA after two washings.
The efficiency of binding was determined by quantification of the residual concentration of peptides in the buffer after overnight incubation with the beads.
Nonspecific binding was determined with 5 μM -vesamicol.
Specific binding was determined by subtracting values of non-specific binding from that of total binding.
After a final wash, cells were fixed in 1% paraformaldehyde and SEB binding was determined by flow cytometry (FACSCalibur, Becton Dickinson, Mountain View, CA).
Non-specific binding was determined with 10 µM MLA.
Non-specific binding was determined with 20 µM unlabeled rosiglitazone.
Nonspecific binding was determined with 10 mM GBR 12935 (Sigma).
Non-specific binding was determined as binding in the presence of 10 mM non-radioactive ouabain.
Specific binding was determined by subtracting non-specific binding density from total binding density.
Peptide binding was determined using ELISA.
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