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When to use GST: When you want easy affinity purification with mild conditions Pull-down assays (GST pull-down is a standard technique) When dimerization is acceptable or desired Quick-and-dirty expression screening When NOT to use GST: When you need monomeric protein When maximum solubility enhancement is critical For crystallization of the passenger protein For any assay where dimerization state matters Thioredoxin (Trx) Size: ~12 kDa (109 residues) Mechanism: Thioredoxin promotes cytoplasmic disulfide bond formation when expressed in appropriate E

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