Placenta · 2016
Don't trust an(t)ybody - Pitfalls during investigation of candidate proteins for methylmercury transport at the placental interface.
СтатьяНазвание и аннотация: Перевод в очереди
Обработка статьи · этапы, ошибки и управление
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Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
Ellinger Isabella, Chatuphonprasert Waranya, Reiter Martin, Voss Agnes, Kemper Jost, Straka Elisabeth, Scheinast Matthias, Zeisler Harald, Salzer Hans, Gundacker Claudia
Аннотация
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.
Предметные рубрики MeSH
Adaptor Proteins, Signal TransducingBiological TransportCarrier ProteinsFemaleFusion Regulatory Protein 1, Heavy ChainHumansLarge Neutral Amino Acid-Transporter 1Methylmercury CompoundsMultidrug Resistance-Associated Protein 2ATP-Binding Cassette, Sub-Family C ProteinsPlacentaPregnancyRNA, Small Interfering