Muller S, Chaix M L, Briand J P, Van Regenmortel M H
Laboratoire d'Immunochimie, Institut de Biologie Moléculaire et Cellulaire, CNRS, Strasbourg, France.
Mol Immunol. 1991 Jul;28(7):763-72. doi: 10.1016/0161-5890(91)90119-5.
Histone antibodies have been obtained by immunizing rabbits with histones H1, H2A, H2B, H3, H4 and triacetylated H4, uncomplexed to RNA. The reactivity of these antibodies was investigated by ELISA using as antigen isolated histones and chromatin as well as thirty-five different synthetic peptides covering the entire sequence of the four core histones, two peptides of H1 and two acetylated peptides of H4. The binding of these antibodies to histones was also measured in immunoblotting and in microcomplement fixation (MCF) tests. In parallel experiments using the same assays the various antigens were tested with antisera raised against histones complexed with RNA. Antibodies induced in the absence of RNA did not react with histones in MCF tests nor with chromatin in ELISA but reacted with the histones in ELISA, although the antibody titers were somewhat lower than in the case of antisera to histone-RNA complexes. Antibodies to RNA-histone complexes reacted with histones in both ELISA and MCF tests. When they were tested with peptide-coated microtiter plates in a direct binding ELISA format, antibodies induced with uncomplexed histones recognized very few fragments which were mainly located in the N- and C-terminal ends of the histones.
通过用未与RNA复合的组蛋白H1、H2A、H2B、H3、H4和三乙酰化H4免疫兔子获得了组蛋白抗体。使用分离的组蛋白和染色质作为抗原，以及覆盖四种核心组蛋白整个序列的35种不同合成肽、两种H1肽和两种H4乙酰化肽，通过ELISA研究了这些抗体的反应性。还通过免疫印迹和微量补体固定（MCF）试验测量了这些抗体与组蛋白的结合。在使用相同检测方法的平行实验中，用针对与RNA复合的组蛋白产生的抗血清检测了各种抗原。在无RNA情况下诱导产生的抗体在MCF试验中不与组蛋白反应，在ELISA中也不与染色质反应，但在ELISA中与组蛋白反应，尽管抗体效价比针对组蛋白-RNA复合物的抗血清略低。针对RNA-组蛋白复合物的抗体在ELISA和MCF试验中均与组蛋白反应。当在直接结合ELISA形式中用肽包被的微量滴定板对它们进行检测时，由未复合组蛋白诱导产生的抗体识别的片段很少，这些片段主要位于组蛋白的N端和C端。
