Should we be cautious on the use of commercially available antibodies to dopamine receptors?
Should we be cautious on the use of commercially available antibodies to dopamine receptors?
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DOI：
10.1007/s00210-008-0384-6
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发表时间：
2009-04-01
影响因子：
3.6
通讯作者：
Sigala, Sandra
中科院分区：
文献类型：
--
作者：
Bodei, Serena;Arrighi, Nicola;Sigala, Sandra
有证据表明，很难获得针对G蛋白偶联受体的特异性抗体，不同的技术困难可能导致产生缺乏特异性的抗体。我们进行了实验，以验证商业上可获得的针对多巴胺（DA）受体hD(1)、hD(4)和hD(5)的抗体的特异性，使用转染方法：我们研究了在HEK 293细胞中选择性地转染各种克隆亚型，是否每种抗体仅在表达其同源受体的细胞中产生条带，而在表达其他DA受体的细胞中不产生条带。我们的研究结果表明，hD(1)和hD(4)受体抗体不仅识别各自的表位，还识别其他DA受体亚型，而对于hD(5)受体检测，我们仅在携带hD(5)转染的HEK 293细胞的通道中观察到信号，尽管缺乏纯度。因此，我们建议谨慎使用市售的DA受体抗体。
Evidence indicate that it is difficult to obtain specific antibodies to G protein-coupled receptors and different technical difficulties may allow the generation of antibodies that lack specificity. We conducted experiments to validate the specificity of commercially available antibodies raised against dopamine (DA) receptors hD(1), hD(4), and hD(5) using a transfection approach: we studied whether, in HEK 293 cells selectively transfected with the various cloned subtypes, each antibody generates bands only in cells expressing its cognate receptor but not in those expressing the other DA receptors. Our results demonstrated that hD(1) and hD(4) receptor antibodies recognize not only their respective epitope, but also other DA receptor subtypes, while for the hD(5) receptor detection, we observed a signal only in the lane loaded with hD(5)-transfected HEK 293 cells, although with a lack of purity. Therefore, we recommend caution on the use of commercially available DA receptor antibodies.