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首页> 外文期刊>BMC Neuroscience >Detection of human and rodent 5-HT 3B receptor subunits by anti-peptide polyclonal antibodies
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Detection of human and rodent 5-HT 3B receptor subunits by anti-peptide polyclonal antibodies

机译：抗肽多克隆抗体检测人和啮齿动物的5-HT 3B受体亚基

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Background The 5-HT3 receptor is a member of a neurotransmitter-gated ion channel family which includes nicotinic acetylcholine, GABAA, and glycine receptors. While antibodies specific for the 5-HT3A receptor subunit are plentiful, and have revealed a wealth of structural and functional information, few antisera exist for the detection of 5-HT3B receptor subunits. Here we describe the generation and characterisation of a rabbit polyclonal antiserum that specifically recognises 5-HT3B receptor subunits Results Immunization of a rabbit with a 20-mer peptide, corresponding to the N-terminus of the human 5-HT3B receptor subunit, generated serum with polyclonal antibodies from which an IgG fraction was purified, yielding pAb77. The antibodies were shown to label 5-HT3B receptor subunits in transfected human embryonic kidney cells and rodent tissues using Western blots. Immunocytochemistry using pAb77 on these cells showed that 5-HT3B receptor subunits do not reach the plasma membrane in the absence of 5-HT3A receptor subunits. Immunohistochemical analysis of rat brain sections showed pAb77 immunoreactivity in distinct populations of cells in the hippocampus. Conclusion We have demonstrated that pAb77 antibodies specifically label native and recombinant 5-HT3B receptor subunits with high affinity and specificity. The antibody was shown to be useful for the determination of both receptor trafficking and also mapping 5-HT3B receptor subunit expression in the CNS.
机译：背景5-HT 3 受体是神经递质门控离子通道家族的成员，该通道包括烟碱乙酰胆碱，GABA A 和甘氨酸受体。虽然对5-HT 3A 受体亚基具有特异性的抗体很多，并且揭示了丰富的结构和功能信息，但用于检测5-HT 3B 的抗血清却很少。受体亚基。在这里，我们描述了特异性识别5-HT 3B 受体亚基的兔多克隆抗血清的产生和表征。结果用20人肽（对应于人5的N端）免疫兔。 -HT 3B 受体亚基产生的多克隆抗体血清可纯化IgG馏分，得到pAb77。使用Western blots显示该抗体标记了转染的人胚胎肾细胞和啮齿动物组织中的5-HT 3B 受体亚基。在这些细胞上使用pAb77进行的免疫细胞化学分析显示，在没有5-HT 3A 受体亚基的情况下，5-HT 3B 受体亚基不会到达质膜。大鼠脑切片的免疫组织化学分析显示，海马中不同细胞群中的pAb77免疫反应性。结论我们已经证明pAb77抗体以高亲和力和特异性特异性标记天然和重组的5-HT 3B 受体亚基。证明该抗体可用于确定受体运输，也可用于绘制CNS中5-HT 3B 受体亚基的表达。

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