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兔抗人CD1d分子α3结构域抗体的制备与鉴定 被引量：1

Preparation and characterization of the rabbit polyclonal antibody against the α3 domain of human CD1d
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摘要 目的:制备兔抗人CD1d分子α3结构域(hCD1d-α3)多克隆抗体。方法:PCR法扩增出人hCD1d-α3编码区基因,将其克隆入原核表达载体pET28中,转化大肠杆菌BL21(DE3),用IPTG诱导重组蛋白的表达,用Ni2+-NTA Agarose亲和层析法纯化重组蛋白,以之为免疫原免疫家兔,制备多克隆抗体,并用ELISA、Western blot及免疫组织化学法检测抗体。结果:成功构建了hCD1d-α3原核表达载体pET28/h CD1d-α3,高效表达并纯化hCD1d-α3蛋白,间接ELISA检测所制备抗体的效价为1∶6400,Western blot显示该抗体能与hCD1d特异结合,免疫组织化学法检测结果表明该抗体识别人小肠组织中的天然hCD1d。结论:通过制备重组hCD1d-α3蛋白为免疫原,免疫家兔,成功地制备了效价高、特异性好的抗hCD1d-α3多克隆抗体。 AIM:To prepare the rabbit antibody against the α3 domain of the human CD1d(hCD1d-α3).METHODS:The gene fragment coding for hCD1d-α3 was amplified by PCR and cloned into prokaryotic expression vector pET28,then expressed in E.coli BL21(DE3).The recombinant protein hCD1d-α3 was purified with Ni2+-NTA agarose column and used as immunogen to immunize the rabbit.The titer and specificity of the anti-hCD1d-α3 antibody from the rabbit were analyzed by ELISA,Western blot and immunohistochemistry,respectively.RESULTS...
作者 陈章权 黄震 梁晓东 何天文 陆田田
机构地区 广东医学院临床免疫学教研室
出处 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2008年第3期250-252,共3页 Chinese Journal of Cellular and Molecular Immunology
基金 广东省中医药局科研基金资助项目(1060043)
关键词 CD1D α3结构域 表达 抗体 制备 CD1d α3 domain expression antibody preparation
分类号 R392.11 [医药卫生—免疫学]
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参考文献10

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共引文献9

同被引文献7

  • 1陈章权,何素辉,陆田田,何天文,何韶衡.兔抗人肥大细胞羧肽酶抗体的制备与鉴定[J].细胞与分子免疫学杂志,2007,23(9):859-861. 被引量：10
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  • 3Haraguchi K, Takahashi T, Nakahara F, et al. CDld expression level in tumor cells is an important determinant for anti-tumor immunity by natural killer T cells[J]. Leuk Lymphoma, 2006, 47(10) : 2218 - 2223.
  • 4Ilyinskii PO, Wang R, Balk SP, et al. CDld mediates T-cell-depend- ent resistance to secondary infection with encephalomyocarditis virus (EMCV) in vitro and immune response to EMCV infection in vivo [J]. J Virol, 2006, 80(14): 7146-7158.
  • 5Chen N, McCarthy C, Drakesmith H, et al. HIV-1 down-regulates the expression of CD1d via Nef[J]. Eur J Immunol, 2006, 36(2) : 278 - 286.
  • 6Fais F, Tenca C, Cimino G, et al. CDld expression on B-precursor acute lymphoblastic leukemia subsets with poor prognosis [ J ]. Leukemia, 2005, 19(4) : 551 -556.
  • 7Kojo S, Tsutsumi A, Goto D, et al. Low expression levels of soluble CDld gene in patients with rheumatoid arthritis [J]. J Rheumatol, 2003, 30(12) :2524 - 2528.

引证文献1

细胞与分子免疫学杂志
2008年 第3期

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