Development of splice variant-specific monoclonal antibodies against human alpha3 integrin
Development of splice variant-specific monoclonal antibodies against human alpha3 integrin
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开发针对人 α3 整合素的剪接变体特异性单克隆抗体
DOI：
10.1089/mab.2015.0053
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Tsuji T
中科院分区：
文献类型：
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作者：
Oku T;Ando Y;Ogura M;Tsuji T
整合素是粘附分子的主要家族，由异源二聚体（α和β亚基）组成。一些报道已经表明在某些整联蛋白亚基的胞质结构域中存在剪接变体。本研究以人脑总RNA为模板，采用RT-PCR方法检测整合素α3剪接变异体（α3A和α3B）的mRNA。α3B变体缺少外显子25编码的序列，似乎是通过选择性剪接产生的。我们建立了小鼠杂交瘤产生的单克隆抗体（这两个都是IgG 1类）的每一个变种的具体。在Western印迹和免疫沉淀实验中，每种抗体对相应的整合素α3变体表现出特异性反应性，表明其是检测相应整合素变体的有用工具。
Integrins are a major family of adhesion molecules, consisting of heterodimers (α and β subunits). Several reports have suggested the presence of splice variants in the cytoplasmic domain of certain integrin subunits. In the present study, we detected mRNA of integrin α3 splice variants (α3A and α3B) by RT-PCR using total RNA from the human brain as a template. The α3B variant lacks the sequence coded by exon 25 and appears to be generated by alternative splicing. We established mouse hybridomas producing monoclonal antibodies (both of which are of IgG1 class) specific for each variant. Each antibody exhibited specific reactivity towards the corresponding integrin α3 variant in Western blotting and immunoprecipitation experiments, suggesting it to be a useful tool for detection of the respective integrin variant.