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Practice question

Question

Which tag is commonly used in TAP for high-purity complex isolation?

Options

Choose one · Correct answer highlighted

Explanation

Tandem Affinity Purification relies on Calmodulin Binding Peptide or CBP as second affinity module for high-purity complex isolation under near-physiological conditions. CBP binds calmodulin resin in calcium-dependent manner after first purification on IgG Sepharose and TEV cleavage. Calcium chelation with EGTA allows gentle elution preserving native complexes for downstream mass spectrometry. TRITC is fluorescent dye, biotin requires streptavidin matrix, and GFP is reporter for localization, not an affinity handle in classic TAP design. CBP provides low background and reversible binding essential for sequential purification strategy maximizing purity.