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使用一种独特的佐剂生成一系列针对天然绿色荧光蛋白的单克隆抗体,用于免疫染色、荧光激活细胞分选、免疫沉淀和染色质免疫沉淀。

Generating a battery of monoclonal antibodies against native green fluorescent protein for immunostaining, FACS, IP, and ChIP using a unique adjuvant.

作者信息

Sanchez Philip, Daniels Karla J, Park Yang-Nim, Soll David R

机构信息

Developmental Studies Hybridoma Bank, The University of Iowa , Iowa City, Iowa.

出版信息

Monoclon Antib Immunodiagn Immunother. 2014 Apr;33(2):80-8. doi: 10.1089/mab.2013.0089.

Abstract

Using a unique, nontoxic adjuvant compound of poly(I:C) and anti-CD40 MAb, a battery of eight mouse monoclonal antibodies was generated against native green fluorescent protein. All were effective to varying degrees for immunostaining paraformaldehyde-fixed cells, six for staining sections of paraffin-embedded tissue, all to varying degrees in fluorescent-activated cell sorting, five for immunoprecipitation, and seven for chromatin immunoprecipitation. None worked in denaturing Western blots since the target was the native GFP protein. Both the hybridomas and antibodies are available at cost through DSHB, a non-profit National Resource created by the National Institutes of Health.

摘要

使用聚肌胞苷酸(poly(I:C))和抗CD40单克隆抗体的独特无毒佐剂化合物,产生了一组针对天然绿色荧光蛋白的八种小鼠单克隆抗体。所有抗体在不同程度上对免疫染色多聚甲醛固定的细胞有效,六种对石蜡包埋组织切片染色有效,在荧光激活细胞分选方面均有不同程度的效果,五种用于免疫沉淀,七种用于染色质免疫沉淀。由于靶标是天然GFP蛋白,因此在变性蛋白质免疫印迹中均无效。杂交瘤和抗体均可通过DSHB按成本价获取,DSHB是美国国立卫生研究院创建的非营利性国家资源。

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