CN200510072500
CN1765927A
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摘要:抗Axin的抗体C8的制备方法,涉及一种抗Axin的抗体C8的制备方法。提供一种效价高、特异性强、能用于进行内源Axin的免疫印迹、免疫沉淀分析的抗Axin的抗体C8制备方法。先用PGR方法扩增出挪威鼠表达Axin 751-832氨基酸序列DNA片段,并连接到原核表达载体pGST parallel中构建重组质粒,再转化到大肠杆菌BL21菌株中,通过IPTG诱导,表达出融合有GST的Axin片断GST-Axin751-832;用谷胱甘肽-琼脂糖树脂纯化该抗原蛋白片断,4次免疫新西兰兔后,采血,纯化抗体血清。能快速大量获得抗原蛋白,抗体纯化方法独特,成本低,过程简单。其抗体效价高,特异性强。能检测到细胞内含量极其稀少的内源Axin。
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Abstract: The preparation method for antibody C8 to anti-Axin with high titer, strong specificity, and fit to synthesis for immunoblotting and deposition, which comprises: with PGR method, amplifying the DNA fragment of Norway mouse expressed Axin 751-832 amino acid sequence, linking into pGST parallel to construct recombinant plasmid; then transforming to strain of BL21 of Escherichia coli; with IPTG inducing, expressing GST-Axin751-832 fused with Axin fragment of GST; with glutathion-agarose resin, purifying the albumen fragment; immuning the New Zealand rabbit for four times; gathering blood, purifying antibody serum. This method can obtain mass antibody albumen rapidly with low cost, and the product can detect much less inner Axin.
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申请人: 厦门大学
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Applicant: UNIV XIAMEN[CN]
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地址: 361005福建省********(隐藏)
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发明(设计)人: 骆晶晶 贺颖 陈明谅 林圣彩
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Inventor: LUO JINGJING HE[CN]
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主分类号: C07K16/18(2006.01)I
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分类号: C07K16/18(2006.01)I C12N15/70(2006.01)I C12P19/34(2006.01)I
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