Detection of immunolabels with multi-isotope imaging mass spectrometry (MIMS).
Detection of immunolabels with multi-isotope imaging mass spectrometry (MIMS).
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DOI：
10.1002/sia.5596
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发表时间：
2014-11-01
期刊：
影响因子：
--
通讯作者：
Lechene C
中科院分区：
文献类型：
--
作者：
Thiery-Lavenant G;Guillermier C;Wang M;Lechene C
我们开发了一种结合使用稳定同位素、MIMS和抗体的方法。我们首先在小鼠的肠道细胞中使用了成熟的抗体，抗肌动蛋白和抗突触素。我们将该方法扩展到免疫金分析，以特异地定位视网膜突触带的主要蛋白质成分Ribeye，或定位含有突触小泡的突触蛋白--突触素。两者均定位于视网膜光感受器细胞的突触前神经末梢。我们的结果表明，通过对Au信号的MIMS分析，我们可以直接识别标记了未扩增的1.4 nm金纳米颗粒的抗体。他们还证明，金纳米颗粒标记的抗体不会稀释用于测量蛋白质周转的15N/14N信号。因此，我们可以同时和直接使用MIMS来测量蛋白质周转和识别细胞类型或特定的蛋白质。
We have developed a method that combines the use of stable isotopes, MIMS and antibody. We began with using well-established antibodies, anti-actin and anti-synaptophysin, in mouse intestinal cells. We extended the method to an immunogold assay to specifically localize Ribeye, a major protein component of retina synaptic ribbons, or to localize a synaptic vesicle-containing protein, synaptophysin. Both are localized in presynaptic nerve terminal of photoreceptors cells in retina. Our results show that by MIMS analysis of the Au signal we can directly identify antibodies tagged with non amplified 1.4 nm gold nanoparticles. They also demonstrate that the gold nanoparticle-tagged antibodies do not dilute the 15N/14N signal used for measuring protein turnover. Thus we can simultaneously and directly use MIMS to measure protein turnover and to identify cell type or specific protein.