不要轻信任何人——胎盘界面甲基汞转运候选蛋白研究中的陷阱。 （注：原英文中an(t)ybody拼写有误，推测正确为anybody）
Don't trust an(t)ybody - Pitfalls during investigation of candidate proteins for methylmercury transport at the placental interface.
作者信息
Ellinger Isabella, Chatuphonprasert Waranya, Reiter Martin, Voss Agnes, Kemper Jost, Straka Elisabeth, Scheinast Matthias, Zeisler Harald, Salzer Hans, Gundacker Claudia
机构信息
Department of Pathophysiology and Allergy Research, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria.
Institute of Medical Genetics, Center of Pathobiochemistry and Genetics, Medical University Vienna, Vienna, Austria.
出版信息
Placenta. 2016 Jul;43:13-6. doi: 10.1016/j.placenta.2016.04.011. Epub 2016 Apr 20.
While investigating placental mercury transport, we validated specificity of commercial antibodies against four candidate transporters (Large neutral amino acids transporter (LAT)1, LAT2, 4F2 cell-surface antigen heavy chain (4F2hc), and multidrug resistance-associated protein (MRP)2) by immunoblotting and small interfering RNA (siRNA)-mediated protein knockdown. An anti-4F2hc- and one anti-LAT1-antibody were specific. Another anti-LAT1-antibody reacted with LAT2. Two anti-LAT2-antibodies detected mainly albumin in placental lysates. A specific anti-MRP2-antibody hardly detected MRP2 in human placentas, contradicting published data. We recommend testing any unknown antibody by western blotting for 1/specificity for the protein of interest using e.g. siRNA knockdown and 2/cross-reactivity with albumin.
在研究胎盘汞转运过程中，我们通过免疫印迹法和小干扰RNA（siRNA）介导的蛋白质敲低，验证了针对四种候选转运蛋白（大中性氨基酸转运蛋白（LAT）1、LAT2、4F2细胞表面抗原重链（4F2hc）和多药耐药相关蛋白（MRP）2）的商业抗体的特异性。一种抗4F2hc抗体和一种抗LAT1抗体具有特异性。另一种抗LAT1抗体与LAT2发生反应。两种抗LAT2抗体在胎盘裂解物中主要检测到白蛋白。一种特异性抗MRP2抗体在人胎盘中几乎检测不到MRP2，这与已发表的数据相矛盾。我们建议通过蛋白质免疫印迹法，利用例如siRNA敲低技术，检测任何未知抗体对目标蛋白的1/特异性，以及2/与白蛋白的交叉反应性。
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