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小鼠主要组织相容性复合体H-2Kk分子的纯化

Purification of the H-2Kk molecule of the murine major histocompatibility complex.

作者信息

Herrmann S H, Mescher M F

出版信息

J Biol Chem. 1979 Sep 25;254(18):8713-6.

PMID:479148
Abstract

The intact H-2Kk antigen has been detergent-solubilized and purified using an immunoabsorbent column prepared from the 11-4.1 monoclonal antibody described by Oi et al. (Oi, V. T., Jones, P. P., Goding, J. Current Topics in Microbiology and Immunology (Melchers, F., Potter, M., and Warner, N. L., eds) Vol. 81, pp. 115-129, Springer-Verlag, New York). The mild conditions used for elution from the column, 0.5% deoxycholate in 10 mM Tris buffer, pH 8, with 0.14 M NaCl, result in recovery of 70 to 100% of the allogeneic serological activity. A murine lymphoma, RDM-4, was found to express high levels of H2-Kk; approximately 2 X 10(6) molecules/cell. Milligram quantities of H-2Kk can be purified readily using these cells.

摘要

完整的H-2Kk抗原已用去污剂溶解,并使用由Oi等人描述的11-4.1单克隆抗体制备的免疫吸附柱进行纯化(Oi, V. T., Jones, P. P., Goding, J.《微生物学与免疫学当前专题》(Melchers, F., Potter, M., and Warner, N. L., 编)第81卷,第115 - 129页,施普林格出版社,纽约)。从柱上洗脱所使用的温和条件,即在pH 8的10 mM Tris缓冲液中加入0.5%脱氧胆酸盐和0.14 M NaCl,可使70%至100%的同种异体血清学活性得以恢复。发现一种鼠淋巴瘤RDM-4表达高水平的H2-Kk;大约每细胞2×10⁶个分子。使用这些细胞可以很容易地纯化出毫克量的H-2Kk。

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