Halikowski M J, Liew C C
Biochem J. 1985 Jan 15;225(2):357-63. doi: 10.1042/bj2250357.
Three monoclonal antibody subclasses (IgG1, IgG2a, and IgM) were raised to the phosphoprotein B2 (Mr 68000, pI6.5-8.2) which has been shown previously to be associated with the nucleosomes of rat liver nuclei. These antibodies do not show any significant cross reactivity with CM-cellulose 'unbound' non-histone chromosomal proteins, bovine serum albumin or histones. Further verification of the specificity of these antibodies to this phosphoprotein was carried out using both 'dot' blot and immunological transfer analysis ('Western blot'). The monoclonal antibodies (IgG1 and IgG2a) could also be used to semi-quantify the phosphoprotein B2 in rat liver nuclei. The high specificity and unlimited availability of this type of probe provides a means to study the role(s) of this phosphoprotein in the overall scheme of actively transcribed chromatin.
针对磷蛋白B2（分子量68000，等电点6.5 - 8.2）制备了三种单克隆抗体亚类（IgG1、IgG2a和IgM），该磷蛋白先前已被证明与大鼠肝细胞核的核小体相关。这些抗体与CM - 纤维素“未结合”的非组蛋白染色体蛋白、牛血清白蛋白或组蛋白均无明显交叉反应。使用“斑点”印迹法和免疫转移分析（“Western印迹法”）对这些抗体针对该磷蛋白的特异性进行了进一步验证。单克隆抗体（IgG1和IgG2a）还可用于半定量大鼠肝细胞核中的磷蛋白B2。这类探针的高特异性和无限可用性为研究该磷蛋白在活跃转录染色质整体机制中的作用提供了一种手段。
