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抗人PD-1高亲和力抗血清的制备及其在PD-1免疫印迹分析中的应用

Preparation of anti-human PD-1 antiserum with high-affinity and its application in PD-1 immunoblotting analysis
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摘要 利用纯化的可溶性人PD-1胞外域(sPD-1)蛋白为免疫原免疫小鼠,获得抗血清.免疫印迹显示该抗血清与原核表达的sPD-1有特异性反应,但与某些菌体蛋白存在交叉反应.经菌体蛋白吸附法处理后,交叉反应明显减弱.以商业化抗人PD-1抗体和小鼠sPD-1抗血清分析经亲和层析富集的活化的Jurkat T细胞表达的PD-1,只有自制的抗血清鉴定到特异反应条带,其表观分子质量为49 ku. The anti-human soluble program death-1 (sPD-1) antiserum was prepared by immunizing mice with the purified recombinant sPD-1. Immunoblotting analysis showed that this antiserum had specific reaction with sPD-1, and had some cross-reactivity with bacterial proteins. This cross-reactivity was significantly decreased after treating the antiserum with whole bacterial lysates. PD-1 expressed by activated Jurkat T cells was enriched by affinity chromatography, and assayed by commercial antibody and the antiserum respectively. Only the antiserum identified the natural PD-1, its apparent molecular weight is about 49 000.
出处 《暨南大学学报(自然科学与医学版)》 CAS CSCD 北大核心 2008年第3期315-319,共5页 Journal of Jinan University(Natural Science & Medicine Edition)
基金 国家自然科学基金项目(30572199、30230350、30371651) 生物化学与分子生物学广东省重点学科资助()
关键词 程序死亡蛋白-1 JURKAT T细胞 抗血清 免疫印迹 programmed death-1 Jurkat T cell antiserum immunoblotting
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