制备一系列针对萤火虫荧光素酶的单克隆抗体，用于斑点印迹分析、蛋白质免疫印迹分析以及对培养细胞和石蜡切片进行免疫染色。

Generating a Battery of Monoclonal Antibodies Against Firefly Luciferase for Dot Blot Analysis, Western Blot Analysis, and Immunostaining of Cells in Culture and Paraffin Sections.

作者信息

Conway Thomas P, Daniels Karla J, Park Yang-Nim, Soll David R

机构信息

Developmental Studies Hybridoma Bank and W.M. Keck Dynamic Image Analysis, Department of Biology, The University of Iowa , Iowa City, Iowa.

出版信息

Monoclon Antib Immunodiagn Immunother. 2018 Feb;37(1):45-51. doi: 10.1089/mab.2017.0057.

DOI:10.1089/mab.2017.0057

PMID:29474160

Abstract

Firefly luciferase (FLuc) is commonly used as a reporter gene PpyLuc1 in bioanalytical assays. We have produced five mouse-derived monoclonal antibodies (mAbs) that recognize FLuc. The mAbs, DSHB-LUC-2, DSHB-LUC-3, DSHB-LUC-9, DSHB-LUC-16, and DSHB-LUC-24, were generated by immunizing mice with purified 6xHIS-tagged FLuc (6xHis-FLuc) in suspension with an adjuvant. All five were validated by dot blots. Four of the mAbs provided strong signals in western blot analysis, and one a weak signal. All five were validated for immunostaining in fixed cell culture. Only one stained cells embedded in paraffin. The five mAbs are available at cost through the Developmental Studies Hybridoma Bank (DSHB), a nonprofit National Resource created by the National Institutes of Health.

摘要

萤火虫荧光素酶（FLuc）在生物分析检测中通常用作报告基因PpyLuc1。我们制备了五种识别FLuc的小鼠源单克隆抗体（mAb）。这些单克隆抗体，即DSHB-LUC-2、DSHB-LUC-3、DSHB-LUC-9、DSHB-LUC-16和DSHB-LUC-24，是通过用纯化的6xHIS标记的FLuc（6xHis-FLuc）与佐剂悬浮免疫小鼠产生的。所有五种抗体均通过斑点印迹法进行了验证。其中四种单克隆抗体在蛋白质印迹分析中提供了强信号，一种提供了弱信号。所有五种抗体均在固定细胞培养中进行了免疫染色验证。只有一种对石蜡包埋的细胞进行了染色。这五种单克隆抗体可通过发育研究杂交瘤库（DSHB）以成本价获得，DSHB是美国国立卫生研究院创建的一个非营利性国家资源库。