Glutathione S-transferase–Mage-a3 (full length) was produced in BL21(DE3) cells by overnight induction at 16ºC with 0.5 mM isopropyl β-d-1-thiogalactopyranoside (IPTG). Protein was purified from bacterial lysates with glutathione Sepharose (GE Amersham) and eluted with 10 mM glutathione. Purified recombinant protein was sent to Cocalico Biologicals Inc. for polyclonal antibody production. Crude antibody serum was affinity-purified against full-length recombinant Mage-a3. Antibody is predicted to recognize Mage-a1/Mage-a2/Mage-a3/Mage-a5/Mage-a6/Mage-a8 proteins but not Mage-a4 and Mage-a10.

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