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目的: 制备高效价、特异性的兔抗人MAGE-n血清。方法: 对原核表达的MAGE-n蛋白进行纯化, 与弗氏佐剂混合免疫新西兰兔制备抗血清, 用硫酸铵盐析法及溴化氰活化的Sephrose4B(CNBr-ActivatedSephrose4B)亲和层析柱纯化抗血清, 以双向免疫扩散实验、ELISA、Westernblot和免疫组化对兔抗人MAGE-n血清进行鉴定。结果: 经免疫得到高效价的兔抗人MAGE-n血清, 抗血清能够与MAGE-n特异性结合, 免疫组化结果表明MAGE-n是一种胞质蛋白。结论: 用MAGE-n蛋白与弗氏佐剂混合免疫兔, 制备得到高效价的抗血清, 纯化后的特异性兔抗人MAGE-n血清, 为进一步研究MAGE-n在各种组织中的表达奠定了基础。
Objective: To prepare high titer and specificity rabbit anti-human MAGE-n serum. Methods: The prokaryotic expression of MAGE-n protein was purified and mixed with Freund’s adjuvant to immunize New Zealand rabbits to prepare antiserum. The antiserum was purified by ammonium sulfate salification and cyanogen bromide-activated Sephrose 4B (CNBr-ActivatedSephrose 4B) affinity chromatography Antiserum, rabbit anti-human MAGE-n serum was identified by two-dimensional immunodiffusion assay, ELISA, Western blot and immunohistochemistry. Results: After high titer of rabbit anti-human MAGE-n serum was obtained by immunization, the antiserum could specifically bind to MAGE-n. Immunohistochemistry showed that MAGE-n was a kind of cytoplasmic protein. Conclusion: MAGE-n protein is mixed with Freund’s adjuvant to immunize rabbits to prepare high-titer antiserum and purified rabbit anti-human MAGE-n serum. To further study the expression of MAGE-n in various tissues Foundation.